Difference between revisions of "Team:Mingdao/Parts"

 
(12 intermediate revisions by 2 users not shown)
Line 1: Line 1:
−
{{:Mingdao_remove_header}}
+
<html>
−
 
+
<head>
−
<!DOCTYPE html>
+
<style>
−
<html class="html">
+
.firstHeading {
−
<head>
+
display: none;
 +
}
 +
#globalWrapper, #content{
 +
background-color: transparent;
 +
border: 0px;
 +
float: left;
 +
        margin:0px;
 +
padding: 0px;
 +
        width:100%;
 +
}
 +
</style>
  
 +
  <script type="text/javascript">
 +
  if(typeof Muse == "undefined") window.Muse = {}; window.Muse.assets = {"required":["jquery-1.8.3.min.js", "museutils.js", "jquery.watch.js", "jquery.musemenu.js", "parts.css"], "outOfDate":[]};
 +
</script>
 +
 
 
   <meta http-equiv="Content-type" content="text/html;charset=UTF-8"/>
 
   <meta http-equiv="Content-type" content="text/html;charset=UTF-8"/>
−
   <meta name="generator" content="7.4.30.244"/>
+
   <meta name="generator" content="2014.0.0.264"/>
−
   <title>BioBrick</title>
+
   <title>Parts</title>
 
   <!-- CSS -->
 
   <!-- CSS -->
−
   <link rel="stylesheet" type="text/css" href="https://2014hs.igem.org/Mingdao_CSS_site_global?action=raw&ctype=text/css"/>
+
   <link rel="stylesheet" type="text/css" href="https://2015.igem.org/Team:Mingdao/CSS-site_global?action=raw&ctype=text/css"/>
−
  <link rel="stylesheet" type="text/css" href="https://2014hs.igem.org/Mingdao_CSS_master_main-template?action=raw&ctype=text/css"/>
+
    <style>
−
  <link rel="stylesheet" type="text/css" href="https://2014hs.igem.org/Mingdao_CSS_bio-brick?action=raw&ctype=text/css" id="pagesheet"/>
+
#u4110
 +
{
 +
border-color: #000000;
 +
background: transparent url("https://static.igem.org/mediawiki/2015/9/96/Mingdao-marmod2.png") no-repeat center top;
 +
background-size: cover;
 +
}
 +
 
 +
#u99
 +
{
 +
border-color: #000000;
 +
background-color: #E7E0D9;
 +
background-color: rgba(117, 76, 36, 0.17);
 +
-pie-background: rgba(117, 76, 36, 0.17);
 +
}
 +
 
 +
#u5104
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u5104:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u5104:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u5104.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1135
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u1135:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u1135:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1135.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1184
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u1184:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u1184:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1184.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1179
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u1179:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u1179:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1179.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1240
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u1240:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u1240:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1240.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1142
 +
{
 +
background-color: transparent;
 +
position: relative;
 +
}
 +
 
 +
#u1142:hover
 +
{
 +
background-color: #DDDDDD;
 +
background-color: rgba(153, 153, 153, 0.33);
 +
-pie-background: rgba(153, 153, 153, 0.33);
 +
}
 +
 
 +
#u1142:active
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u1142.MuseMenuActive
 +
{
 +
background-color: #AE7566;
 +
background-color: rgba(121, 26, 0, 0.6);
 +
-pie-background: rgba(121, 26, 0, 0.6);
 +
}
 +
 
 +
#u5107-4_clip,#u1137-4_clip,#u1185-4_clip,#u1181-6_clip,#u1242-6_clip,#u1145-6_clip
 +
{
 +
overflow: hidden;
 +
vertical-align: top;
 +
}
 +
 
 +
#u5107-4_img,#u1137-4_img,#u1185-4_img,#u1181-6_img,#u1242-6_img,#u1145-6_img
 +
{
 +
display: block;
 +
}
 +
 
 +
.MenuItem /* unifiedNavBar */
 +
{
 +
cursor: pointer;
 +
}
 +
 
 +
#u100
 +
{
 +
border-color: #000000;
 +
background-color: #E9D9C7;
 +
background-color: rgba(0,0,0,0);
 +
position: relative;
 +
-pie-background: rgba(0,0,0,0);
 +
}
 +
 
 +
#u4110_align_to_page,#u99_align_to_page,#menuu1133,#u5103,#u5107-4,#u1134,#u1137-4,#u1183,#u1185-4,#u1148,#u1181-6,#u1239,#u1242-6,#u1141,#u1145-6,#u8651
 +
{
 +
position: relative;
 +
}
 +
 
 +
#u1246,#u1247,#u1248,#u1249,#u5113
 +
{
 +
border-style: solid;
 +
border-color: #7F3400;
 +
background-color: transparent;
 +
position: relative;
 +
border-width: 0px 0px 0px 2px;
 +
}
 +
.version.parts /* version checker */
 +
{
 +
color: #0000EB;
 +
background-color: #1F158E;
 +
}
 +
 
 +
.html
 +
{
 +
background: #7F7F7F url("https://static.igem.org/mediawiki/2015/8/8f/Mingdao-bg.gif") repeat left top scroll;
 +
}
 +
#all{
 +
      width:100%;
 +
      background: #FFFFFF url("https://static.igem.org/mediawiki/2015/8/8f/Mingdao-bg.gif") repeat-y center top;
 +
}
 +
 
 +
#page
 +
{
 +
z-index: 1;
 +
width: 1280px;
 +
min-height: 12242.750000000005px;
 +
border-style: none;
 +
border-color: #000000;
 +
padding-bottom: 0px;
 +
margin-left: auto;
 +
margin-right: auto;
 +
background: #FFFFFF url("https://static.igem.org/mediawiki/2015/8/8f/Mingdao-bg.gif") repeat-y center top;
 +
}
 +
 
 +
#page_position_content
 +
{
 +
padding-bottom: 48px;
 +
}
 +
 
 +
#pu4110
 +
{
 +
width: 0.01px;
 +
margin-left: -100px;
 +
}
 +
 
 +
#u4110
 +
{
 +
z-index: 2;
 +
padding-bottom: 0px;
 +
}
 +
 
 +
#u4110_align_to_page
 +
{
 +
margin-left: auto;
 +
margin-right: auto;
 +
width: 1280px;
 +
left: -100px;
 +
padding-top: 640px;
 +
padding-bottom: 5px;
 +
}
 +
 
 +
#u4202
 +
{
 +
z-index: 55;
 +
width: 696px;
 +
left: 392px;
 +
position: relative;
 +
}
 +
 
 +
#pu4204-4
 +
{
 +
width: 0.01px;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4204-4
 +
{
 +
z-index: 56;
 +
display: block;
 +
vertical-align: top;
 +
position: relative;
 +
margin-right: -10000px;
 +
left: 46px;
 +
}
 +
 
 +
#u4205-4
 +
{
 +
z-index: 60;
 +
width: 70px;
 +
min-height: 29px;
 +
font-size: 72px;
 +
line-height: 86px;
 +
color: #F15A24;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 44px;
 +
}
 +
 
 +
#u4203-4
 +
{
 +
z-index: 64;
 +
width: 24px;
 +
min-height: 29px;
 +
font-size: 72px;
 +
line-height: 86px;
 +
color: #F15A24;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 44px;
 +
left: 672px;
 +
}
 +
 
 +
#u4206-4
 +
{
 +
z-index: 68;
 +
width: 977px;
 +
font-size: 20px;
 +
line-height: 32px;
 +
color: #FFFFFF;
 +
left: 261px;
 +
margin-top: 276px;
 +
position: relative;
 +
}
 +
 
 +
#u4207-4
 +
{
 +
z-index: 72;
 +
display: block;
 +
vertical-align: top;
 +
left: 541px;
 +
margin-top: 48px;
 +
position: relative;
 +
}
 +
 
 +
#u4208-10
 +
{
 +
z-index: 76;
 +
width: 1079px;
 +
left: 215px;
 +
top: -12px;
 +
margin-bottom: -12px;
 +
position: relative;
 +
}
 +
 
 +
#u4110-bw
 +
{
 +
z-index: 2;
 +
height: 1645px;
 +
}
 +
 
 +
#u99
 +
{
 +
z-index: 3;
 +
padding-bottom: 14px;
 +
}
 +
 
 +
#u99_align_to_page
 +
{
 +
margin-left: auto;
 +
margin-right: auto;
 +
width: 1280px;
 +
left: -100px;
 +
}
 +
 
 +
#menuu1133
 +
{
 +
z-index: 5;
 +
width: 829px;
 +
margin-right: -10000px;
 +
margin-top: 14px;
 +
left: 531px;
 +
}
 +
 
 +
#u5103
 +
{
 +
width: 136px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u5104
 +
{
 +
width: 136px;
 +
padding-bottom: 40px;
 +
}
 +
 
 +
#u5104:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u5104:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1134
 +
{
 +
width: 136px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
left: 138px;
 +
}
 +
 
 +
#u1135
 +
{
 +
width: 136px;
 +
padding-bottom: 40px;
 +
}
 +
 
 +
#u5104.MuseMenuActive,#u1135:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1135:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1183
 +
{
 +
width: 136px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
left: 276px;
 +
}
 +
 
 +
#u1184
 +
{
 +
width: 136px;
 +
padding-bottom: 40px;
 +
}
 +
 
 +
#u1135.MuseMenuActive,#u1184:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1184:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u5107-4_clip,#u1137-4_clip,#u1185-4_clip
 +
{
 +
width: 136px;
 +
height: 32px;
 +
}
 +
 
 +
#u5107-4,#u1137-4,#u1185-4
 +
{
 +
margin-right: -10000px;
 +
top: 20px;
 +
}
 +
 
 +
#u1148
 +
{
 +
width: 137px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
left: 414px;
 +
}
 +
 
 +
#u1179
 +
{
 +
width: 137px;
 +
padding-bottom: 8px;
 +
}
 +
 
 +
#u1184.MuseMenuActive,#u1179:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1179:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1239
 +
{
 +
width: 137px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
left: 553px;
 +
}
 +
 
 +
#u1240
 +
{
 +
width: 137px;
 +
padding-bottom: 8px;
 +
}
 +
 
 +
#u1179.MuseMenuActive,#u1240:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1240:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1141
 +
{
 +
width: 137px;
 +
min-height: 72px;
 +
margin-right: -10000px;
 +
left: 692px;
 +
}
 +
 
 +
#u1142
 +
{
 +
width: 137px;
 +
padding-bottom: 8px;
 +
}
 +
 
 +
#u1240.MuseMenuActive,#u1142:hover
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1142:active
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1142.MuseMenuActive
 +
{
 +
margin: 0px;
 +
}
 +
 
 +
#u1181-6_clip,#u1242-6_clip,#u1145-6_clip
 +
{
 +
width: 137px;
 +
height: 64px;
 +
}
 +
 
 +
#u1181-6,#u1242-6,#u1145-6
 +
{
 +
margin-right: -10000px;
 +
top: 4px;
 +
}
 +
 
 +
#u99-bw
 +
{
 +
z-index: 3;
 +
margin-top: 208px;
 +
height: 86px;
 +
}
 +
 
 +
#u100
 +
{
 +
z-index: 4;
 +
width: 333px;
 +
padding-bottom: 8px;
 +
margin-right: -10000px;
 +
margin-top: 142px;
 +
left: 186px;
 +
}
 +
 
 +
#u8651
 +
{
 +
z-index: 53;
 +
width: 317px;
 +
margin-right: -10000px;
 +
margin-top: -19px;
 +
left: 12px;
 +
}
 +
 
 +
#u1246
 +
{
 +
z-index: 48;
 +
width: 23px;
 +
height: 58px;
 +
margin-right: -10000px;
 +
margin-top: 232px;
 +
left: 668px;
 +
}
 +
 
 +
#u1247
 +
{
 +
z-index: 49;
 +
width: 23px;
 +
height: 58px;
 +
margin-right: -10000px;
 +
margin-top: 232px;
 +
left: 805px;
 +
}
 +
 
 +
#u1248
 +
{
 +
z-index: 50;
 +
width: 23px;
 +
height: 58px;
 +
margin-right: -10000px;
 +
margin-top: 232px;
 +
left: 1082px;
 +
}
 +
 
 +
#u1249
 +
{
 +
z-index: 51;
 +
width: 23px;
 +
height: 58px;
 +
margin-right: -10000px;
 +
margin-top: 232px;
 +
left: 1221px;
 +
}
 +
 
 +
#u5113
 +
{
 +
z-index: 52;
 +
width: 24px;
 +
height: 58px;
 +
margin-right: -10000px;
 +
margin-top: 232px;
 +
left: 943px;
 +
}
 +
 
 +
#gene
 +
{
 +
position: relative;
 +
width: 22px;
 +
margin-right: -10000px;
 +
margin-top: 1259px;
 +
left: 646px;
 +
}
 +
 
 +
#menuu6069
 +
{
 +
z-index: 494;
 +
width: 186px;
 +
position: fixed;
 +
top: 133px;
 +
left: -163px;
 +
}
 +
 
 +
#u6140
 +
{
 +
width: 186px;
 +
position: relative;
 +
}
 +
 
 +
#u6143
 +
{
 +
width: 186px;
 +
min-height: 22px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 24px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6143:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6143:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6143.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6087
 +
{
 +
width: 186px;
 +
min-height: 22px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 24px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6087:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6087:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6087.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6197
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6197:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6197:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6197.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6108
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6108:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6108:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6108.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6236
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6236:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6236:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6236.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6229
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6229:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6229:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6229.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6222
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6222:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6222:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6222.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6136
 +
{
 +
width: 186px;
 +
min-height: 22px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 24px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6136:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6136:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6136.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6144-4,#u6090-4,#u6139-4
 +
{
 +
width: 186px;
 +
min-height: 22px;
 +
color: #FFFFFF;
 +
font-size: 18px;
 +
line-height: 22px;
 +
text-align: right;
 +
position: relative;
 +
margin-right: -10000px;
 +
top: 12px;
 +
}
 +
 
 +
#u6169
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 29px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6169:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6169:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6169.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6199-4,#u6110-4,#u6239-4,#u6232-4,#u6225-4,#u6172-4
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
color: #FFFFFF;
 +
font-size: 14px;
 +
text-align: right;
 +
position: relative;
 +
margin-right: -10000px;
 +
top: 14px;
 +
}
 +
 
 +
#u6176
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 30px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6176:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6176:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6176.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6084,#u6196,#u6105,#u6235,#u6228,#u6221,#u6133,#u6168,#u6175,#u6154
 +
{
 +
width: 186px;
 +
margin-top: 2px;
 +
position: relative;
 +
}
 +
 
 +
#u6157
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
background-color: #FFE7B5;
 +
background-color: rgba(255, 213, 121, 0.55);
 +
padding-bottom: 30px;
 +
position: relative;
 +
margin-right: -10000px;
 +
-pie-background: rgba(255, 213, 121, 0.55);
 +
}
 +
 
 +
#u6157:hover
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6157:active
 +
{
 +
background-color: #ADADAD;
 +
background-color: rgba(107, 107, 107, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(107, 107, 107, 0.55);
 +
}
 +
 
 +
#u6157.MuseMenuActive
 +
{
 +
background-color: #C6C6C6;
 +
background-color: rgba(153, 153, 153, 0.55);
 +
margin: 0px -10000px 0px 0px;
 +
-pie-background: rgba(153, 153, 153, 0.55);
 +
}
 +
 
 +
#u6179-4,#u6158-4
 +
{
 +
width: 186px;
 +
min-height: 17px;
 +
color: #FFFFFF;
 +
font-size: 14px;
 +
text-align: right;
 +
position: relative;
 +
margin-right: -10000px;
 +
top: 15px;
 +
}
 +
 
 +
#u6144-2,#u6090-2,#u6199-2,#u6110-2,#u6239-2,#u6232-2,#u6225-2,#u6139-2,#u6172-2,#u6179-2,#u6158-2
 +
{
 +
padding-right: 8px;
 +
}
 +
 
 +
.MenuItem /* unifiedNavBar */
 +
{
 +
cursor: pointer;
 +
}
 +
 
 +
#biobrick
 +
{
 +
margin-left: 496px;
 +
margin-top: 12px;
 +
position: relative;
 +
}
 +
 
 +
#u4209-4
 +
{
 +
z-index: 86;
 +
display: block;
 +
vertical-align: top;
 +
margin-left: 441px;
 +
top: -3px;
 +
margin-bottom: -3px;
 +
position: relative;
 +
}
 +
 
 +
#u4210-4
 +
{
 +
z-index: 90;
 +
width: 1116px;
 +
font-size: 20px;
 +
line-height: 24px;
 +
color: #FFFFFF;
 +
margin-left: 98px;
 +
margin-top: 16px;
 +
position: relative;
 +
}
 +
 
 +
#sr-psb1c3
 +
{
 +
margin-left: 557px;
 +
margin-top: 49px;
 +
position: relative;
 +
}
 +
 
 +
#u4211-21
 +
{
 +
z-index: 94;
 +
width: 1097px;
 +
margin-left: 106px;
 +
margin-top: 9px;
 +
position: relative;
 +
}
 +
 
 +
#u4254
 +
{
 +
z-index: 268;
 +
width: 1105px;
 +
margin-left: 88px;
 +
position: relative;
 +
}
 +
 
 +
#srpk-psb1c3
 +
{
 +
margin-left: 530px;
 +
margin-top: 48px;
 +
position: relative;
 +
}
 +
 
 +
#u4230-24
 +
{
 +
z-index: 115;
 +
width: 1097px;
 +
margin-left: 84px;
 +
margin-top: 7px;
 +
position: relative;
 +
}
 +
 
 +
#u4260
 +
{
 +
z-index: 270;
 +
width: 1026px;
 +
margin-left: 127px;
 +
margin-top: 7px;
 +
position: relative;
 +
}
 +
 
 +
#u4243-4
 +
{
 +
z-index: 190;
 +
width: 1168px;
 +
min-height: 20px;
 +
font-size: 20px;
 +
line-height: 24px;
 +
color: #FFFFFF;
 +
margin-left: 56px;
 +
margin-top: 25px;
 +
position: relative;
 +
}
 +
 
 +
#gst-sr-psb1c3
 +
{
 +
margin-left: 485px;
 +
margin-top: 105px;
 +
position: relative;
 +
}
 +
 
 +
#pu4234-24
 +
{
 +
width: 0.01px;
 +
margin-left: 74px;
 +
margin-top: 23px;
 +
}
 +
 
 +
#u4234-24
 +
{
 +
z-index: 139;
 +
width: 1097px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4266
 +
{
 +
z-index: 272;
 +
width: 996px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 279px;
 +
left: 68px;
 +
}
 +
 
 +
#u4242-4
 +
{
 +
z-index: 186;
 +
width: 1165px;
 +
color: #FFFFFF;
 +
font-size: 20px;
 +
line-height: 24px;
 +
font-family: Arial, Helvetica Neue, Helvetica, sans-serif;
 +
margin-left: 57px;
 +
margin-top: 21px;
 +
position: relative;
 +
}
 +
 
 +
#srpk-his-psb1c3
 +
{
 +
margin-left: 512px;
 +
margin-top: 20px;
 +
position: relative;
 +
}
 +
 
 +
#u4238-23
 +
{
 +
z-index: 163;
 +
width: 1097px;
 +
margin-left: 91px;
 +
margin-top: 11px;
 +
position: relative;
 +
}
 +
 
 +
#u4211-2,#u4230-2,#u4234-2,#u4238-2
 +
{
 +
font-size: 28px;
 +
text-align: center;
 +
line-height: 34px;
 +
color: #FBB03B;
 +
}
 +
 
 +
#u4208,#u4208-5,#u4211-3,#u4211-10,#u4211-16,#u4230-3,#u4230-17,#u4234-3,#u4234-17,#u4238-3,#u4238-17
 +
{
 +
color: #FBB03B;
 +
}
 +
 
 +
#u4230-12,#u4230-21,#u4234-12,#u4234-21,#u4238-12,#u4238-21
 +
{
 +
font-size: 20px;
 +
line-height: 24px;
 +
color: #FBB03B;
 +
}
 +
 
 +
#u4272
 +
{
 +
z-index: 274;
 +
width: 903px;
 +
margin-left: 189px;
 +
margin-top: 13px;
 +
position: relative;
 +
}
 +
 
 +
#data
 +
{
 +
margin-left: 574px;
 +
margin-top: 43px;
 +
position: relative;
 +
}
 +
 
 +
#u4244-4
 +
{
 +
z-index: 194;
 +
display: block;
 +
vertical-align: top;
 +
margin-left: 456px;
 +
margin-top: 8px;
 +
position: relative;
 +
}
 +
 
 +
#u4245-4
 +
{
 +
z-index: 198;
 +
width: 634px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 323px;
 +
margin-top: 9px;
 +
position: relative;
 +
}
 +
 
 +
#pu4278
 +
{
 +
width: 0.01px;
 +
margin-left: 415px;
 +
margin-top: 43px;
 +
}
 +
 
 +
#u4278
 +
{
 +
z-index: 276;
 +
width: 200px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4278_clip
 +
{
 +
overflow: hidden;
 +
width: 200px;
 +
height: 232px;
 +
}
 +
 
 +
#u4278_img
 +
{
 +
margin: -6px -5px;
 +
}
 +
 
 +
#u4246-14
 +
{
 +
z-index: 202;
 +
width: 309px;
 +
font-size: 20px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 26px;
 +
left: 245px;
 +
}
 +
 
 +
#u4247-4
 +
{
 +
z-index: 216;
 +
width: 616px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 326px;
 +
margin-top: 42px;
 +
position: relative;
 +
}
 +
 
 +
#pu4284
 +
{
 +
width: 0.01px;
 +
margin-left: 294px;
 +
margin-top: 51px;
 +
}
 +
 
 +
#u4284
 +
{
 +
z-index: 278;
 +
width: 342px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4284_clip
 +
{
 +
overflow: hidden;
 +
width: 342px;
 +
height: 237px;
 +
}
 +
 
 +
#u4284_img
 +
{
 +
margin: -10px -9px -14px -11px;
 +
}
 +
 
 +
#u4248-19
 +
{
 +
z-index: 220;
 +
width: 548px;
 +
position: relative;
 +
margin-right: -10000px;
 +
left: 367px;
 +
}
 +
 
 +
#u4249-4
 +
{
 +
z-index: 239;
 +
width: 1021px;
 +
font-size: 20px;
 +
line-height: 24px;
 +
color: #FFFFFF;
 +
margin-left: 149px;
 +
margin-top: 7px;
 +
position: relative;
 +
}
 +
 
 +
#u4250-4
 +
{
 +
z-index: 243;
 +
width: 612px;
 +
font-size: 25px;
 +
line-height: 38px;
 +
color: #FFFFFF;
 +
margin-left: 334px;
 +
margin-top: 61px;
 +
position: relative;
 +
}
 +
 
 +
#pu4290
 +
{
 +
width: 0.01px;
 +
margin-left: 235px;
 +
margin-top: 52px;
 +
}
 +
 
 +
#u4290
 +
{
 +
z-index: 280;
 +
width: 143px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4290_clip
 +
{
 +
overflow: hidden;
 +
width: 143px;
 +
height: 263px;
 +
}
 +
 
 +
#u4290_img
 +
{
 +
margin: -10px -9px;
 +
}
 +
 
 +
#u4296
 +
{
 +
z-index: 282;
 +
width: 242px;
 +
position: relative;
 +
margin-right: -10000px;
 +
left: 192px;
 +
}
 +
 
 +
#u4296_clip
 +
{
 +
overflow: hidden;
 +
width: 242px;
 +
height: 263px;
 +
}
 +
 
 +
#u4296_img
 +
{
 +
margin: -12px -10px -13px -9px;
 +
}
 +
 
 +
#u4251-8
 +
{
 +
z-index: 247;
 +
width: 429px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 33px;
 +
left: 478px;
 +
}
 +
 
 +
#u4252-4
 +
{
 +
z-index: 255;
 +
width: 788px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 246px;
 +
margin-top: 32px;
 +
position: relative;
 +
}
 +
 
 +
#pu4305
 +
{
 +
width: 0.01px;
 +
margin-left: 125px;
 +
margin-top: 31px;
 +
}
 +
 
 +
#u4305
 +
{
 +
z-index: 284;
 +
width: 538px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4305_clip
 +
{
 +
overflow: hidden;
 +
width: 538px;
 +
height: 266px;
 +
}
 +
 
 +
#u4305_img
 +
{
 +
margin: -7px -7px -9px;
 +
}
 +
 
 +
#u4253-9
 +
{
 +
z-index: 259;
 +
width: 365px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 13px;
 +
left: 588px;
 +
}
 +
 
 +
#u4311-4
 +
{
 +
z-index: 286;
 +
width: 1015px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 133px;
 +
margin-top: 61px;
 +
position: relative;
 +
}
 +
 
 +
#pu4313
 +
{
 +
width: 0.01px;
 +
margin-left: 158px;
 +
margin-top: 66px;
 +
}
 +
 
 +
#u4313
 +
{
 +
z-index: 313;
 +
width: 244px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 11px;
 +
}
 +
 
 +
#u4319
 +
{
 +
z-index: 315;
 +
width: 146px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 11px;
 +
left: 294px;
 +
}
 +
 
 +
#u4312-23
 +
{
 +
z-index: 290;
 +
width: 494px;
 +
font-size: 20px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
position: relative;
 +
margin-right: -10000px;
 +
left: 472px;
 +
}
 +
 
 +
#u4325-4
 +
{
 +
z-index: 317;
 +
width: 631px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 325px;
 +
margin-top: 78px;
 +
position: relative;
 +
}
 +
 
 +
#pu4327
 +
{
 +
width: 0.01px;
 +
margin-left: 325px;
 +
margin-top: 38px;
 +
}
 +
 
 +
#u4327
 +
{
 +
z-index: 333;
 +
width: 193px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4326-12
 +
{
 +
z-index: 321;
 +
width: 534px;
 +
font-size: 20px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 41px;
 +
left: 225px;
 +
}
 +
 
 +
#u4333-4
 +
{
 +
z-index: 335;
 +
width: 548px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 366px;
 +
margin-top: 52px;
 +
position: relative;
 +
}
 +
 
 +
#u4334-16
 +
{
 +
z-index: 339;
 +
width: 351px;
 +
margin-left: 466px;
 +
margin-top: 43px;
 +
position: relative;
 +
}
 +
 
 +
#u4334-13
 +
{
 +
font-size: 20px;
 +
text-align: center;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
}
 +
 
 +
#vector
 +
{
 +
margin-left: 508px;
 +
margin-top: 10px;
 +
position: relative;
 +
}
 +
 
 +
#u4335-4
 +
{
 +
z-index: 355;
 +
display: block;
 +
vertical-align: top;
 +
margin-left: 441px;
 +
top: -1px;
 +
margin-bottom: -1px;
 +
position: relative;
 +
}
 +
 
 +
#u4336-4
 +
{
 +
z-index: 359;
 +
width: 839px;
 +
font-size: 20px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
margin-left: 220px;
 +
margin-top: 14px;
 +
position: relative;
 +
}
 +
 
 +
#sr-pgex-2t
 +
{
 +
margin-left: 552px;
 +
margin-top: 10px;
 +
position: relative;
 +
}
 +
 
 +
#u4337-9
 +
{
 +
z-index: 363;
 +
width: 971px;
 +
margin-left: 156px;
 +
margin-top: 9px;
 +
position: relative;
 +
}
 +
 
 +
#u4338
 +
{
 +
z-index: 372;
 +
width: 490px;
 +
margin-left: 386px;
 +
top: -6px;
 +
margin-bottom: -6px;
 +
position: relative;
 +
}
 +
 
 +
#u4254_img,#u4338_img
 +
{
 +
padding-right: 1px;
 +
}
 +
 
 +
#srpk-pet-29b
 +
{
 +
margin-left: 532px;
 +
margin-top: 30px;
 +
position: relative;
 +
}
 +
 
 +
#u4344-8
 +
{
 +
z-index: 374;
 +
width: 971px;
 +
margin-left: 145px;
 +
margin-top: 11px;
 +
position: relative;
 +
}
 +
 
 +
#u4337-2,#u4344-2
 +
{
 +
font-size: 25px;
 +
text-align: center;
 +
line-height: 38px;
 +
color: #FBB03B;
 +
}
 +
 
 +
#u4345
 +
{
 +
z-index: 382;
 +
width: 498px;
 +
margin-left: 382px;
 +
margin-top: 12px;
 +
position: relative;
 +
}
 +
 
 +
#data2
 +
{
 +
margin-left: 554px;
 +
margin-top: 10px;
 +
position: relative;
 +
}
 +
 
 +
#pu4351-5
 +
{
 +
width: 0.01px;
 +
margin-left: 184px;
 +
margin-top: 12px;
 +
}
 +
 
 +
#u4351-5
 +
{
 +
z-index: 384;
 +
width: 159px;
 +
position: relative;
 +
margin-right: -10000px;
 +
left: 376px;
 +
}
 +
 
 +
#u4351-2
 +
{
 +
font-size: 30px;
 +
text-align: center;
 +
line-height: 45px;
 +
letter-spacing: 3px;
 +
color: #FBB03B;
 +
}
 +
 
 +
#u4352-6
 +
{
 +
z-index: 389;
 +
width: 893px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 46px;
 +
}
 +
 
 +
#u4352-2
 +
{
 +
font-size: 25px;
 +
text-align: center;
 +
line-height: 38px;
 +
color: #B0DCFF;
 +
}
 +
 
 +
#u4208-3,#u4208-4,#u4208-7,#u4248-2,#u4248-4,#u4248-6,#u4248-8,#u4248-10,#u4248-12,#u4248-14,#u4248-16,#u4251-2,#u4251-3,#u4251-5,#u4253-2,#u4253-4,#u4253-6,#u4334-2,#u4334-4,#u4334-6,#u4334-8,#u4334-9,#u4337-4,#u4337-6,#u4344-4,#u4344-6,#u4352-4
 +
{
 +
font-size: 20px;
 +
line-height: 30px;
 +
color: #FFFFFF;
 +
}
 +
 
 +
#u4353-4
 +
{
 +
z-index: 395;
 +
width: 287px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #91DDFF;
 +
margin-left: 487px;
 +
margin-top: 31px;
 +
position: relative;
 +
}
 +
 
 +
#pu4354
 +
{
 +
width: 0.01px;
 +
margin-left: 221px;
 +
margin-top: 31px;
 +
}
 +
 
 +
#u4354
 +
{
 +
z-index: 399;
 +
width: 266px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4360-16
 +
{
 +
z-index: 401;
 +
width: 615px;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 50px;
 +
left: 299px;
 +
}
 +
 
 +
#u4360-2,#u4360-11
 +
{
 +
font-size: 23px;
 +
line-height: 28px;
 +
color: #91DDFF;
 +
}
 +
 
 +
#u4362-4
 +
{
 +
z-index: 417;
 +
width: 188px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #91DDFF;
 +
margin-left: 537px;
 +
margin-top: 48px;
 +
position: relative;
 +
}
 +
 
 +
#u4363-4
 +
{
 +
z-index: 421;
 +
width: 383px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #F4FF97;
 +
margin-left: 439px;
 +
margin-top: 30px;
 +
position: relative;
 +
}
 +
 
 +
#pu4366
 +
{
 +
width: 0.01px;
 +
margin-left: 262px;
 +
margin-top: 49px;
 +
}
 +
 
 +
#u4366
 +
{
 +
z-index: 452;
 +
width: 325px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4364-27
 +
{
 +
z-index: 425;
 +
display: block;
 +
vertical-align: top;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 6px;
 +
left: 369px;
 +
}
 +
 
 +
#u4372-4
 +
{
 +
z-index: 454;
 +
width: 347px;
 +
font-size: 25px;
 +
line-height: 30px;
 +
color: #F4FF97;
 +
margin-left: 457px;
 +
margin-top: 50px;
 +
position: relative;
 +
}
 +
 
 +
#pu4374
 +
{
 +
width: 0.01px;
 +
margin-left: 268px;
 +
margin-top: 26px;
 +
}
 +
 
 +
#u4374
 +
{
 +
z-index: 480;
 +
width: 325px;
 +
position: relative;
 +
margin-right: -10000px;
 +
}
 +
 
 +
#u4373-22
 +
{
 +
z-index: 458;
 +
display: block;
 +
vertical-align: top;
 +
position: relative;
 +
margin-right: -10000px;
 +
margin-top: 60px;
 +
left: 369px;
 +
}
 +
 
 +
#u4377-12
 +
{
 +
z-index: 482;
 +
width: 830px;
 +
margin-left: 216px;
 +
margin-top: 31px;
 +
position: relative;
 +
}
 +
 
 +
#u4377-2,#u4377-7
 +
{
 +
font-size: 25px;
 +
text-align: center;
 +
line-height: 30px;
 +
color: #91DDFF;
 +
}
 +
 
 +
#u4211-7,#u4211-9,#u4211-11,#u4211-13,#u4211-15,#u4211-18,#u4230-8,#u4230-10,#u4230-14,#u4230-16,#u4230-19,#u4234-8,#u4234-10,#u4234-14,#u4234-16,#u4234-19,#u4238-8,#u4238-10,#u4238-14,#u4238-16,#u4238-19,#u4360-4,#u4360-6,#u4360-8,#u4360-9,#u4360-13,#u4377-4,#u4377-5,#u4377-9
 +
{
 +
font-size: 20px;
 +
line-height: 24px;
 +
color: #FFFFFF;
 +
}
 +
 
 +
body
 +
{
 +
position: relative;
 +
min-width: 1280px;
 +
}
 +
 
 +
#page .verticalspacer
 +
{
 +
clear: both;
 +
}
 +
  </style>
 
   <!-- Other scripts -->
 
   <!-- Other scripts -->
 
   <script type="text/javascript">
 
   <script type="text/javascript">
 
   document.documentElement.className += ' js';
 
   document.documentElement.className += ' js';
 
</script>
 
</script>
−
  </head>
+
  <!-- JS includes -->
−
<body>
+
  <!--[if lt IE 9]>
 +
  <script src="https://2015.igem.org/Team:Mingdao/Javascript-html5shiv?
 +
action=raw&ctype=text/javascript" type="text/javascript"></script>
 +
  <![endif]-->
 +
    <!--custom head HTML-->
 +
  <style>
 +
  #menuu6069{
  
 +
      transition-property: left;
 +
      transition-duration: 0.2s;
 +
      transition-timing-function: ease;
 +
 +
      -webkit-transition-property: left;
 +
      -webkit-transition-duration: 0.2s;
 +
      -webkit-transition-timing-function: ease;
 +
  }
 +
  #menuu6069:hover{
 +
      left:0px;
 +
  }
 +
</style>
 +
</head>
 +
<body>
 +
<div id="all">
 
   <div class="clearfix" id="page"><!-- column -->
 
   <div class="clearfix" id="page"><!-- column -->
 
   <div class="position_content" id="page_position_content">
 
   <div class="position_content" id="page_position_content">
−
     <div class="browser_width colelem" id="u1673"><!-- group -->
+
     <div class="clearfix colelem" id="pu4110"><!-- group -->
−
     <div class="clearfix" id="u1673_align_to_page">
+
     <div class="browser_width grpelem" id="u4110-bw">
−
       <div class="clip_frame grpelem" id="u1674"><!-- image -->
+
       <div id="u4110"><!-- column -->
−
      <img class="block" id="u1674_img" src="https://static.igem.org/mediawiki/2014hs/2/2b/Mingdaologo.png" alt="" width="235" height="68"/>
+
      <div class="clearfix" id="u4110_align_to_page">
 +
        <div class="clearfix colelem" id="u4202"><!-- group -->
 +
        <div class="clearfix grpelem" id="pu4204-4"><!-- group -->
 +
          <img class="grpelem" id="u4204-4" alt="Gene Block" width="618" height="121" src="https://static.igem.org/mediawiki/2015/6/60/Mingdao-u4204-4.png"/><!-- rasterized frame -->
 +
          <div class="clearfix grpelem" id="u4205-4"><!-- content -->
 +
          <p>*</p>
 +
          </div>
 +
        </div>
 +
        <div class="clearfix grpelem" id="u4203-4"><!-- content -->
 +
          <p>*</p>
 +
        </div>
 +
        </div>
 +
        <div class="clearfix colelem" id="u4206-4"><!-- content -->
 +
        <p>In our project, we cloned two genes, SR and SRPK from the materials kindly gave by Dr. Woan&#45;Yuh Tarn at Academia Sinica in Taiwan. Phosphoserine and arginine (highest nitrogen contents among 20 amino acids) play important roles in fire retardance. We found a novel fire retardant protein, SR protein, through protein database mining. SR proteins are post&#45;translationally phosphorylated by SR protein kinase (SRPK). We co&#45;transformed E. coli BL21 with two vectors simultaneously to express phosphorylated SR proteins.</p>
 +
        </div>
 +
        <img class="colelem" id="u4207-4" alt="Gene" width="398" height="110" src="https://static.igem.org/mediawiki/2015/5/53/Mingdao-u4207-4.png"/><!-- rasterized frame -->
 +
        <div class="clearfix colelem" id="u4208-10"><!-- content -->
 +
        <p id="u4208-3"><span id="u4208">Serine/arginine&#45;rich splicing factor 1 (SRSF1) </span>is a protein found on chromosome 17 in humans. SRSF1 is also known as alternative splicing factor 1 (ASF1), pre&#45;mRNA&#45;splicing factor SF2 (SF2), which is involved in alternative splicing, mRNA nuclear export and translation. It contains two functional domains. One in the N&#45;terminus is RNA recognition motifs which interact with RNA and splicing factors. The other in the C&#45;terminus is rich in serine and arginine residues (SR domain) and regulate the protein activity through phosphorylation by SR protein kinase.</p>
 +
        <p id="u4208-4">&nbsp;</p>
 +
        <p id="u4208-7"><span id="u4208-5">SRSF protein kinase 1 (SRPK1) </span>is a serine/arginine&#45;rich protein specific kinase, which phosphorylates SR proteins on serine residues in SR domain. It plays an important role in the regulation of SR protein activity. Disorders in SRPK1 may be observed in certain cancer cells.</p>
 +
        <p>&nbsp;</p>
 +
        </div>
 +
      </div>
 
       </div>
 
       </div>
−
       <img class="grpelem" id="u6162-4" src="https://static.igem.org/mediawiki/2014hs/1/1e/U6162-4.png" alt="Odor&#45; Let it Die" width="175" height="42"/><!-- rasterized frame -->
+
    </div>
−
      <div class="clip_frame grpelem" id="u1676"><!-- image -->
+
    <div class="browser_width grpelem" id="u99-bw">
−
      <img class="block" id="u1676_img" src="https://static.igem.org/mediawiki/2014hs/2/29/Igem_official_logo.png" alt="" width="88" height="67"/>
+
       <div id="u99"><!-- group -->
 +
      <div class="clearfix" id="u99_align_to_page">
 +
        <nav class="MenuBar clearfix grpelem" id="menuu1133"><!-- horizontal box -->
 +
        <div class="MenuItemContainer clearfix grpelem" id="u5103"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu rgba-background clearfix colelem" id="u5104" href="https://2015.igem.org/Team:Mingdao"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u5107-4"><!-- rasterized frame --><div id="u5107-4_clip"><img id="u5107-4_img" alt="Home" width="136" height="32" src="https://static.igem.org/mediawiki/2015/9/9d/Mingdao-u5107-4.png"/></div></div></a>
 +
        </div>
 +
        <div class="MenuItemContainer clearfix grpelem" id="u1134"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu rgba-background clearfix colelem" id="u1135" href="https://2015.igem.org/Team:Mingdao/Team"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u1137-4"><!-- rasterized frame --><div id="u1137-4_clip"><img id="u1137-4_img" alt="Team" width="136" height="32" src="https://static.igem.org/mediawiki/2015/1/16/Mingdao-u1137-4.png"/></div></div></a>
 +
        </div>
 +
        <div class="MenuItemContainer clearfix grpelem" id="u1183"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu rgba-background clearfix colelem" id="u1184" href="https://2015.igem.org/Team:Mingdao/Project"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u1185-4"><!-- rasterized frame --><div id="u1185-4_clip"><img id="u1185-4_img" alt="Project" width="136" height="32" src="https://static.igem.org/mediawiki/2015/2/2d/Mingdao-u1185-4.png"/></div></div></a>
 +
        </div>
 +
        <div class="MenuItemContainer clearfix grpelem" id="u1148"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu MuseMenuActive rgba-background clearfix colelem" id="u1179" href="https://2015.igem.org/Team:Mingdao/Parts"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u1181-6"><!-- rasterized frame --><div id="u1181-6_clip"><img id="u1181-6_img" alt="Gene
 +
Block" width="137" height="64" src="https://static.igem.org/mediawiki/2015/0/06/Mingdao-u1181-6.png"/></div></div></a>
 +
        </div>
 +
        <div class="MenuItemContainer clearfix grpelem" id="u1239"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu rgba-background clearfix colelem" id="u1240" href="https://2015.igem.org/Team:Mingdao/Practices"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u1242-6"><!-- rasterized frame --><div id="u1242-6_clip"><img id="u1242-6_img" alt="Human
 +
Practice" width="137" height="64" src="https://static.igem.org/mediawiki/2015/d/d2/Mingdao-u1242-6.png"/></div></div></a>
 +
        </div>
 +
        <div class="MenuItemContainer clearfix grpelem" id="u1141"><!-- vertical box -->
 +
          <a class="nonblock nontext MenuItem MenuItemWithSubMenu rgba-background clearfix colelem" id="u1142" href="https://2015.igem.org/Team:Mingdao/Lab"><!-- horizontal box --><div class="MenuItemLabel NoWrap grpelem" id="u1145-6"><!-- rasterized frame --><div id="u1145-6_clip"><img id="u1145-6_img" alt="Lab
 +
Stuff" width="137" height="64" src="https://static.igem.org/mediawiki/2015/8/84/Mingdao-u1145-6.png"/></div></div></a>
 +
        </div>
 +
        </nav>
 +
      </div>
 
       </div>
 
       </div>
−
      <a class="anchor_item grpelem" id="top"></a>
 
 
     </div>
 
     </div>
 +
    <div class="clearfix grpelem" id="u100"><!-- group -->
 +
      <div class="clip_frame grpelem" id="u8651"><!-- image -->
 +
      <object type="application/x-shockwave-flash" height=300 width=400 data="https://static.igem.org/mediawiki/2015/6/6b/Mingdao-alllogo.swf" style="position:relative; left:-60px; top:-20px;">
 +
<param name="movie" value="https://static.igem.org/mediawiki/2015/6/6b/Mingdao-alllogo.swf" />
 +
<param name="quality" value="high" />
 +
<param name="" value="Exactfit"/>
 +
<param name="wmode" value="transparent">
 +
</object>
 +
      </div>
 +
    </div>
 +
    <div class="grpelem" id="u1246"><!-- simple frame --></div>
 +
    <div class="grpelem" id="u1247"><!-- simple frame --></div>
 +
    <div class="grpelem" id="u1248"><!-- simple frame --></div>
 +
    <div class="grpelem" id="u1249"><!-- simple frame --></div>
 +
    <div class="grpelem" id="u5113"><!-- simple frame --></div>
 +
    <a class="anchor_item grpelem" id="gene"></a>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pu1682-3"><!-- group -->
+
     <nav class="MenuBar clearfix pinned-colelem" id="menuu6069"><!-- vertical box -->
−
     <div class="clearfix browser_width grpelem" id="u1682-3"><!-- content -->
+
    <div class="MenuItemContainer clearfix colelem" id="u6140"><!-- horizontal box -->
−
       <p>&nbsp;</p>
+
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6143" href="Parts#gene"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6144-4"><!-- content --><p id="u6144-2">Gene&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 1</p></div></a>
 +
    </div>
 +
     <div class="MenuItemContainer clearfix colelem" id="u6084"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6087" href="Parts#biobrick"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6090-4"><!-- content --><p id="u6090-2">BioBrick&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6196"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6197" href="Parts#sr-psb1c3"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6199-4"><!-- content --><p id="u6199-2">SR/pSB1C3&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6105"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6108" href="Parts#srpk-psb1c3"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6110-4"><!-- content --><p id="u6110-2">&nbsp;SRPK/pSB1C3&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6235"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6236" href="Parts#gst-sr-psb1c3"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6239-4"><!-- content --><p id="u6239-2">&nbsp;GST&#45;SR/pSB1C3&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6228"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6229" href="Parts#srpk-his-psb1c3"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6232-4"><!-- content --><p id="u6232-2">&nbsp;SRPK&#45;His/pSB1C3&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6221"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6222" href="Parts#data"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6225-4"><!-- content --><p id="u6225-2">&nbsp;DATA&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6133"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6136" href="Parts#vector"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6139-4"><!-- content --><p id="u6139-2">Vector&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 3</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6168"><!-- horizontal box -->
 +
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6169" href="Parts#sr-pgex-2t"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6172-4"><!-- content --><p id="u6172-2">SR/pGEX&#45;2T&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 +
    </div>
 +
    <div class="MenuItemContainer clearfix colelem" id="u6175"><!-- horizontal box -->
 +
       <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6176" href="Parts#srpk-pet-29b"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6179-4"><!-- content --><p id="u6179-2">SRPK/pET&#45;29b&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
 
     </div>
 
     </div>
−
     <ul class="MenuBar clearfix grpelem" id="menuu468"><!-- horizontal box -->
+
     <div class="MenuItemContainer clearfix colelem" id="u6154"><!-- horizontal box -->
−
       <li class="MenuItemContainer clearfix grpelem" id="u616"><!-- vertical box -->
+
       <a class="nonblock nontext MenuItem MenuItemWithSubMenu anim_swing rgba-background clearfix grpelem" id="u6157" href="Parts#data2"><!-- horizontal box --><div class="MenuItemLabel clearfix grpelem" id="u6158-4"><!-- content --><p id="u6158-2">DATA&nbsp;&nbsp;&nbsp;&nbsp; ●</p></div></a>
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u617" href="index.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u619-4"><!-- content --><p>Home</p></div></a>
+
    </div>   </nav>
−
      </li>
+
    <a class="anchor_item colelem" id="biobrick"></a>
−
      <li class="MenuItemContainer clearfix grpelem" id="u623"><!-- vertical box -->
+
    <img class="colelem" id="u4209-4" alt="BioBrick" width="398" height="110" src="https://static.igem.org/mediawiki/2015/2/23/Mingdao-u4209-4.png"/><!-- rasterized frame -->
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u624" href="team.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u625-4"><!-- content --><p>Team</p></div></a>
+
    <div class="clearfix colelem" id="u4210-4"><!-- content -->
−
      </li>
+
    <p>We cloned the genes of SR and SRPK as well as the composite DNA fragments of Ptac&#45;GST&#45;SR and Pt7&#45;SRPK&#45;His onto the standard backbone of pSB1C3 biobrick.</p>
−
      <li class="MenuItemContainer clearfix grpelem" id="u630"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u631" href="project.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u632-4"><!-- content --><p>Project</p></div></a>
+
−
      </li>
+
−
      <li class="MenuItemContainer clearfix grpelem" id="u637"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu MuseMenuActive clearfix colelem" id="u638" href="bio-brick.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u639-4"><!-- content --><p>BioBrick</p></div></a>
+
−
      </li>
+
−
      <li class="MenuItemContainer clearfix grpelem" id="u651"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u652" href="modeling.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u653-4"><!-- content --><p>Modeling</p></div></a>
+
−
      </li>
+
−
      <li class="MenuItemContainer clearfix grpelem" id="u5718"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u5721" href="safety.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u5722-4"><!-- content --><p>Safety</p></div></a>
+
−
      </li>
+
−
      <li class="MenuItemContainer clearfix grpelem" id="u644"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u645" href="human-practices.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u646-4"><!-- content --><p>Human Practices</p></div></a>
+
−
      </li>
+
−
      <li class="MenuItemContainer clearfix grpelem" id="u671"><!-- vertical box -->
+
−
      <a class="nonblock nontext MenuItem MenuItemWithSubMenu clearfix colelem" id="u674" href="media.html"><!-- horizontal box --><div class="MenuItemLabel NoWrap clearfix grpelem" id="u676-4"><!-- content --><p>Media</p></div></a>
+
−
      </li>
+
−
    </ul>
+
−
    <div class="browser_width grpelem" id="u1681"><!-- simple frame --></div>
+
 
     </div>
 
     </div>
−
     <div class="mse_pre_init" id="u2764"><!-- simple frame --></div>
+
     <a class="anchor_item colelem" id="sr-psb1c3"></a>
−
     <div class="clearfix colelem" id="pu3615"><!-- group -->
+
     <div class="clearfix colelem" id="u4211-21"><!-- content -->
−
     <div class="browser_width grpelem" id="u3615"><!-- simple frame --></div>
+
     <p id="u4211-2">SR/pSB1C3</p>
−
     <img class="grpelem" id="u2740-4" src="https://static.igem.org/mediawiki/2014hs/0/0f/U2740-4.png" alt="Bio&#45;Bricks" width="396" height="60"/><!-- rasterized frame -->
+
     <p id="u4211-7"><span id="u4211-3">Part No.:</span>&nbsp;<a class="nonblock" href="http://parts.igem.org/Part:BBa_K1608000">BBa_K1608000</a></p>
 +
    <p id="u4211-9">We designed two primers to amplify the SR genes by PCR, followed by XbaI and PstI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation.</p>
 +
    <p id="u4211-11"><span id="u4211-10">Primers:</span></p>
 +
    <p id="u4211-13">1. SR&#45;XbaI&#45;F: 5'&#45; AAAAGGTCTAGATGTCGGGAGGTGGTGTGATTCGTG &#45;3'</p>
 +
    <p id="u4211-15">2. SR&#45;SpeI&#45;PstI&#45;R: 5'&#45;AAGAAACTGCAGCGGCCGCTACTAGTATTATGTACGAGAGCGAGA &#45;3'</p>
 +
    <p id="u4211-18"><span id="u4211-16">PCR product size:</span>&nbsp;~785 bp</p>
 +
    <p>&nbsp;</p>
 
     </div>
 
     </div>
−
     <div class="shadow Ribbon-Large clearfix browser_width colelem" id="u3442-4"><!-- content -->
+
     <div class="clip_frame colelem" id="u4254"><!-- image -->
−
     <h1 id="u3442-2">Biobrick Map&#45; Click link to go to biobrick description</h1>
+
     <img class="block" id="u4254_img" src="https://static.igem.org/mediawiki/2015/f/fe/Mingdao-t1-1.jpeg" alt="" width="1104" height="500"/>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="u6226-16"><!-- content -->
+
    <a class="anchor_item colelem" id="srpk-psb1c3"></a>
−
     <p>Part Registr<span id="u6226-2">y</span>&nbsp;Name<span id="u6226-4">&nbsp;</span>Attribute<span id="u6226-6">&nbsp;</span>Part (E&#45;X&#45;Part&#45;S&#45;P) <span id="u6226-8">&nbsp;</span>Part Length<span id="u6226-10">&nbsp;</span>Gene<span id="u6226-12">&nbsp;</span>&nbsp;Gene Length</p>
+
     <div class="clearfix colelem" id="u4230-24"><!-- content -->
 +
     <p id="u4230-2">SRPK/pSB1C3</p>
 +
    <p id="u4230-8"><span id="u4230-3">Part No.:</span>&nbsp;<a class="nonblock" href="http://parts.igem.org/Part:BBa_K1608001">BBa_K1608001</a></p>
 +
    <p id="u4230-10">We designed two primers to amplify SRPK1 gene by PCR, followed by EcoRI and SpeI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. However, there’re two PstI sites on the sequence of SRPK1 gene. We’re trying to mutate them to fit the standard biobrick parts.</p>
 +
    <p id="u4230-12">Primers:</p>
 +
    <p id="u4230-14">1. SRPK1&#45;EcoRI&#45;XbaI&#45;F: 5'&#45; AAAAGAATTCGCGGCCGCTTCTAGATGGAGCGGAAAGTGCTTGCGCTCCAG &#45;3'</p>
 +
    <p id="u4230-16">2. SRPK1&#45;SpeI&#45;R: 5'&#45; GGGGGGACTAGTATTAGGAGTTAAGCCAAGGGTG &#45;3'</p>
 +
    <p id="u4230-19"><span id="u4230-17">PCR product size:</span>&nbsp;~1995 bp</p>
 +
    <p id="u4230-21">Plasmid map:</p>
 +
    <p>&nbsp;</p>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="u6225"><!-- group -->
+
     <div class="clip_frame colelem" id="u4260"><!-- image -->
−
     <div class="clearfix grpelem" id="u6224"><!-- column -->
+
     <img class="block" id="u4260_img" src="https://static.igem.org/mediawiki/2015/6/6d/Mingdao-t1-2.jpeg" alt="" width="1026" height="651"/>
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3569" href="bio-brick.html#bb01"><!-- image --><img class="block" id="u3569_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    </div>
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3575" href="bio-brick.html#bb02"><!-- image --><img class="block" id="u3575_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    <div class="clearfix colelem" id="u4243-4"><!-- content -->
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3578" href="bio-brick.html#bb03"><!-- image --><img class="block" id="u3578_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    <p>In order to express SR and SRPK gene more conveniently, we made two composite biobricks with promoter, RBS and a fusion tag.</p>
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3581" href="bio-brick.html#bb004"><!-- image --><img class="block" id="u3581_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    </div>
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3584" href="bio-brick.html#bb05"><!-- image --><img class="block" id="u3584_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    <a class="anchor_item colelem" id="gst-sr-psb1c3"></a>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3587" href="bio-brick.html#bb06"><!-- image --><img class="block" id="u3587_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    <div class="clearfix colelem" id="pu4234-24"><!-- group -->
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u3590" href="bio-brick.html#bb07"><!-- image --><img class="block" id="u3590_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
    <div class="clearfix grpelem" id="u4234-24"><!-- content -->
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3593" href="bio-brick.html#bb08"><!-- image --><img class="block" id="u3593_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
      <p id="u4234-2">GST&#45;SR/pSB1C3</p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3611" href="bio-brick.html#bb09"><!-- image --><img class="block" id="u3611_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-8"><span id="u4234-3">Part No.:</span>&nbsp;<a class="nonblock" href="http://parts.igem.org/Part:BBa_K1608002">BBa_K1608002</a></p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3596" href="bio-brick.html#bb10"><!-- image --><img class="block" id="u3596_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-10">We designed two primers to amplify Ptac&#45;GST&#45;SR DNA fragment by PCR from SR/pGEX&#45;2T, followed by XbaI and PstI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. The SR gene is fused with GST tag and driven by Ptac promoter along with a lac operator downstream.</p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3599" href="bio-brick.html#bb11"><!-- image --><img class="block" id="u3599_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-12">Primers:</p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3602" href="bio-brick.html#bb12"><!-- image --><img class="block" id="u3602_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-14">1. Ptac (pGEX)&#45;XbaI&#45;F: 5'&#45; GAGGGGTCTAGAGTTTTTGCGCCGACATCATAAC &#45;3'</p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3605" href="bio-brick.html#bb13"><!-- image --><img class="block" id="u3605_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-16">2. SR&#45;SpeI&#45;PstI&#45;R: 5'&#45; AAGAAACTGCAGCGGCCGCTACTAGTATTATGTACGAGAGCGAGA &#45;3'</p>
−
       <a class="nonblock nontext anim_swing pinned-colelem" id="u3608" href="bio-brick.html#bb14"><!-- image --><img class="block" id="u3608_img" src="https://static.igem.org/mediawiki/2014hs/6/6e/1402683735_40_hyperlink.png" alt="" width="18" height="18"/></a>
+
       <p id="u4234-19"><span id="u4234-17">PCR product size:</span>&nbsp;~1572 bp</p>
 +
       <p id="u4234-21">Plasmid map:</p>
 +
       <p>&nbsp;</p>
 
     </div>
 
     </div>
−
     <div class="clip_frame grpelem" id="u2915"><!-- image -->
+
     <div class="clip_frame grpelem" id="u4266"><!-- image -->
−
       <img class="block" id="u2915_img" src="https://static.igem.org/mediawiki/2014hs/4/42/Chart.jpg" alt="" width="1015" height="382"/>
+
       <img class="block" id="u4266_img" src="https://static.igem.org/mediawiki/2015/e/e9/Mingdao-t1-3.jpeg" alt="" width="996" height="617"/>
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pu3197-4"><!-- group -->
+
     <div class="clearfix colelem" id="u4242-4"><!-- content -->
−
    <div class="shadow Ribbon-Large clearfix browser_width grpelem" id="u3197-4"><!-- content -->
+
     <p>In order to express SR and SRPK gene more conveniently, we made two composite biobricks with promoter, RBS and a fusion tag.</p>
−
      <h1 id="u3197-2">Description</h1>
+
−
     </div>
+
−
    <a class="anchor_item grpelem" id="biobrickdesign"></a>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pu3380"><!-- group -->
+
    <a class="anchor_item colelem" id="srpk-his-psb1c3"></a>
−
     <div class="clearfix grpelem" id="u3380"><!-- column -->
+
     <div class="clearfix colelem" id="u4238-23"><!-- content -->
−
      <div class="clearfix colelem" id="u3315-4"><!-- content -->
+
     <p id="u4238-2">SRPK&#45;His/pSB1C3</p>
−
      <p>Biobrick Design</p>
+
    <p id="u4238-8"><span id="u4238-3">Part No.:</span>&nbsp;<a class="nonblock" href="http://parts.igem.org/Part:BBa_K1608003">BBa_K1608003</a></p>
 +
    <p id="u4238-10">&nbsp;We designed two primers to amplify Pt7&#45;SRPK1&#45;His DNA fragment by PCR from SRPK/pET&#45;29b, followed by EcoRI and SpeI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. However, there’re two PstI sites one XbaI site on the amplified sequence. We’re trying to mutate them to fit the standard biobrick parts. The SRPK1 gene is fused with 6XHis tag and driven by Pt7 promoter along with a lac operator downstream.</p>
 +
    <p id="u4238-12">Primers:</p>
 +
    <p id="u4238-14">1. Pt7(pET)&#45;EcoRI&#45;XbaI&#45;F: 5'&#45; AAAAAAGAATTCGCGGCCGCTTCTAGAGCACGATGCGTCCGGCGTA &#45;3'</p>
 +
    <p id="u4238-16">2. His (pET)&#45;SpeI&#45;R: 5'&#45; AGGGAAACTAGTATCAGTGGTGGTGGTGGTGGT &#45;3'</p>
 +
    <p id="u4238-19"><span id="u4238-17">PCR product size: </span>~2294 bp</p>
 +
    <p id="u4238-21">Plasmid map:</p>
 +
    </div>
 +
    <div class="clip_frame colelem" id="u4272"><!-- image -->
 +
    <img class="block" id="u4272_img" src="https://static.igem.org/mediawiki/2015/4/45/Mingdao-t1-4.jpeg" alt="" width="903" height="571"/>
 +
    </div>
 +
    <a class="anchor_item colelem" id="data"></a>
 +
    <img class="colelem" id="u4244-4" alt="Data" width="368" height="80" src="https://static.igem.org/mediawiki/2015/0/0f/Mingdao-u4244-4.png"/><!-- rasterized frame -->
 +
    <div class="clearfix colelem" id="u4245-4"><!-- content -->
 +
    <p>(1) PCR of SR, SRPK, Ptac&#45;GST&#45;SR and Pt7&#45;SRPK&#45;His</p>
 +
    </div>
 +
    <div class="clearfix colelem" id="pu4278"><!-- group -->
 +
    <div class="clip_frame clearfix grpelem" id="u4278"><!-- image -->
 +
      <div id="u4278_clip">
 +
      <img class="position_content" id="u4278_img" src="https://static.igem.org/mediawiki/2015/f/f1/Mingdao-t1-5.jpeg" alt="" width="209" height="244"/>
 
       </div>
 
       </div>
−
      <div class="colelem" id="u3316"><!-- simple frame --></div>
 
 
     </div>
 
     </div>
−
     <a class="anchor_item grpelem" id="bb01"></a>
+
     <div class="clearfix grpelem" id="u4246-14"><!-- content -->
 +
      <p>Lanes:</p>
 +
      <p>1. 1kb DNA marker</p>
 +
      <p>2. Pt7&#45;SRPK&#45;His: 2294 bp</p>
 +
      <p>3. Ptac&#45;GST&#45;SR: 1572 bp</p>
 +
      <p>4. SR: 785 bp</p>
 +
      <p>5. SRPK: 1995 bp</p>
 +
    </div>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="u3466"><!-- column -->
+
     <div class="clearfix colelem" id="u4247-4"><!-- content -->
−
    <div class="clearfix colelem" id="pu3317-21"><!-- group -->
+
    <p>(2) Restriction enzyme digestion of inserts and vectors</p>
−
      <div class="clearfix grpelem" id="u3317-21"><!-- content -->
+
    </div>
−
      <p id="u3317-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256001">pSB1C3&#45;Plac&#45;NB (Part: BBa_K1256001)</a></p>
+
    <div class="clearfix colelem" id="pu4284"><!-- group -->
−
      <p>&nbsp;</p>
+
    <div class="clip_frame clearfix grpelem" id="u4284"><!-- image -->
−
      <p id="u3317-8"><span id="u3317-6">BENEFIT: </span>cloning genes with NheI &amp; BamHI sites, easily exchange RFP gene</p>
+
      <div id="u4284_clip">
−
      <p id="u3317-11"><span id="u3317-9">BIOBRICK LIGATION</span>:</p>
+
       <img class="position_content" id="u4284_img" src="https://static.igem.org/mediawiki/2015/1/12/Mingdao-t1-6.png" alt="" width="362" height="261"/>
−
      <p id="u3317-13">&#45; “X&#45;P” ligated to “S&#45;P” ( X, S are compatible): standard part</p>
+
−
      <p id="u3317-15">&#45; “E&#45;N” ligated to “E&#45;X” (N, X are compatible): to exchange front part (Fig 1. A)</p>
+
−
      <p id="u3317-17">&#45; “N&#45;P” ligated to “S&#45;P” (N, S are compatible): to exchange back part (Fig 1. B)</p>
+
−
      <p>&nbsp;</p>
+
−
      <p>&nbsp;</p>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3320"><!-- image -->
+
−
       <img class="block" id="u3320_img" src="https://static.igem.org/mediawiki/2014hs/7/70/Psb1c3-plac-nb.png" alt="" width="507" height="174"/>
+
 
       </div>
 
       </div>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="u3352-9"><!-- content -->
+
     <div class="clearfix grpelem" id="u4248-19"><!-- content -->
−
       <p>Fig 1. BioBrick “part in part”&nbsp; (image below)</p>
+
       <p id="u4248-2">Lanes:</p>
 +
      <p id="u4248-4">1. 1kb DNA marker</p>
 +
      <p id="u4248-6">2. Pt7&#45;SRPK&#45;His: EcoRI + SpeI → 2294 bp</p>
 +
      <p id="u4248-8">3. SRPK: EcoRI + SpeI →1995 bp</p>
 +
      <p id="u4248-10">4. pSB1C3: EcoRI + SpeI → 2047 + 1092 bp*</p>
 +
      <p id="u4248-12">5. Ptac&#45;GST&#45;SR: XbaI + PstI→ 1572 bp</p>
 +
      <p id="u4248-14">6. SR: XbaI + PstI → 785 bp</p>
 +
      <p id="u4248-16">7. pSB1C3: XbaI + PstI → 2044 + 1095 bp*</p>
 
       <p>&nbsp;</p>
 
       <p>&nbsp;</p>
−
      <p id="u3352-7">(A<span id="u3352-5">)</span>(B)</p>
 
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pu3358"><!-- group -->
+
    </div>
−
      <div class="clip_frame grpelem" id="u3358"><!-- image -->
+
    <div class="clearfix colelem" id="u4249-4"><!-- content -->
−
       <img class="block" id="u3358_img" src="https://static.igem.org/mediawiki/2014hs/f/f5/Biobrick_fig_b.png" alt="" width="297" height="305"/>
+
     <p>*Bands shifted were caused by the fluorescent DNA loading dye. We confirmed with the specialist and sequenced the plasmid. The plasmids were correct but it bothers us for a long time. Just be very careful to use a ready&#45;to&#45;use fluorescent DNA loading dye.</p>
 +
    </div>
 +
    <div class="clearfix colelem" id="u4250-4"><!-- content -->
 +
    <p>(3)Colony PCR for SRPK/pSB1C3 and SR/pSB1C3</p>
 +
    </div>
 +
    <div class="clearfix colelem" id="pu4290"><!-- group -->
 +
    <div class="clip_frame clearfix grpelem" id="u4290"><!-- image -->
 +
      <div id="u4290_clip">
 +
       <img class="position_content" id="u4290_img" src="https://static.igem.org/mediawiki/2015/8/87/Mingdao-t1-7.jpeg" alt="" width="161" height="283"/>
 
       </div>
 
       </div>
−
      <div class="clip_frame grpelem" id="u3353"><!-- image -->
+
    </div>
−
       <img class="block" id="u3353_img" src="https://static.igem.org/mediawiki/2014hs/0/0d/Biobrick_fig_a.png" alt="" width="297" height="305"/>
+
    <div class="clip_frame clearfix grpelem" id="u4296"><!-- image -->
 +
      <div id="u4296_clip">
 +
       <img class="position_content" id="u4296_img" src="https://static.igem.org/mediawiki/2015/c/c5/Mingdao-t1-8.jpeg" alt="" width="261" height="288"/>
 
       </div>
 
       </div>
 
     </div>
 
     </div>
−
    </div>
+
     <div class="clearfix grpelem" id="u4251-8"><!-- content -->
−
    <a class="anchor_item colelem" id="bb02"></a>
+
       <p id="u4251-2">Left panel: Colony PCR with primers of SRPK1&#45;EcoRI&#45;XbaI&#45;F and VR to check SRPK/pSB1C3 (2162 bp)</p>
−
    <div class="clearfix colelem" id="u3493"><!-- group -->
+
       <p id="u4251-3">&nbsp;</p>
−
     <div class="clearfix grpelem" id="u3366-16"><!-- content -->
+
       <p id="u4251-5">Right panel: Colony PCR with primers of SR&#45;XbaI&#45;F + VR to check SR/pSB1C3 (941 bp)</p>
−
       <p id="u3366-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256002">pSB1C3&#45;Plac&#45;NB&#45;Cm&#45;MN (BBa_K1256002)</a></p>
+
−
       <p id="u3366-5">&nbsp;</p>
+
−
       <p id="u3366-8"><span id="u3366-6">BENEFIT:</span>&nbsp;cloning genes with MfeI &amp; NsiI sites under CmR cassette</p>
+
−
      <p id="u3366-10">BIOBRICK LIGATION:</p>
+
−
      <p id="u3366-12">&#45; “M&#45;Ns” ligated to “E&#45;P” ( M, E are compatible; Ns, P are compatible)</p>
+
 
       <p>&nbsp;</p>
 
       <p>&nbsp;</p>
−
      <p>&nbsp;</p>
 
−
    </div>
 
−
    <div class="clip_frame grpelem" id="u3487"><!-- image -->
 
−
      <img class="block" id="u3487_img" src="https://static.igem.org/mediawiki/2014hs/0/02/Psb1c3-plac-nb-cm-mn.png" alt="" width="519" height="193"/>
 
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
    <a class="anchor_item colelem" id="bb03"></a>
+
     <div class="clearfix colelem" id="u4252-4"><!-- content -->
−
     <div class="clearfix colelem" id="pattack"><!-- group -->
+
    <p>(4)Colony PCR for Ptac&#45;GST&#45;SR/pSB1C3 and Pt7&#45;SRPK&#45;His/pSB1C3</p>
−
    <a class="anchor_item grpelem" id="attack"></a>
+
−
    <div class="clearfix grpelem" id="u3512"><!-- group -->
+
−
      <div class="clearfix grpelem" id="u3374-20"><!-- content -->
+
−
      <p id="u3374-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256003">pSB1C3&#45;Plac&#45;SS&#45;NB (Part: BBa_K1256003)</a></p>
+
−
      <p id="u3374-5">&nbsp;</p>
+
−
      <p id="u3374-8"><span id="u3374-6">Secretion signal (SS):</span>&nbsp;OmpA SS from E. coli</p>
+
−
      <p id="u3374-11"><span id="u3374-9">BENEFIT:</span>&nbsp;cloning genes with NheI &amp; BamHI sites, easily exchange RFP gene</p>
+
−
      <p id="u3374-13">BIOBRICK LIGATION:</p>
+
−
      <p id="u3374-15">&#45; “X&#45;P” ligated to “S&#45;P” or “N&#45;P” ligated to “E&#45;X” (X, S, N are compatible)</p>
+
−
      <p>&nbsp;</p>
+
−
      <p>&nbsp;</p>
+
−
      <p>&nbsp;</p>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3507"><!-- image -->
+
−
      <img class="block" id="u3507_img" src="https://static.igem.org/mediawiki/2014hs/0/05/Psb1c3-plac-ss-nb.png" alt="" width="504" height="173"/>
+
−
      </div>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pbb004"><!-- group -->
+
     <div class="clearfix colelem" id="pu4305"><!-- group -->
−
    <a class="anchor_item grpelem" id="bb004"></a>
+
     <div class="clip_frame clearfix grpelem" id="u4305"><!-- image -->
−
     <div class="clearfix grpelem" id="u3381"><!-- column -->
+
       <div id="u4305_clip">
−
       <div class="clearfix colelem" id="u3383-4"><!-- content -->
+
      <img class="position_content" id="u4305_img" src="https://static.igem.org/mediawiki/2015/a/a7/Mingdao-t1-9.jpeg" alt="" width="552" height="282"/>
−
      <p>Attack</p>
+
 
       </div>
 
       </div>
−
      <div class="colelem" id="u3382"><!-- simple frame --></div>
 
 
     </div>
 
     </div>
−
    </div>
+
     <div class="clearfix grpelem" id="u4253-9"><!-- content -->
−
    <div class="clearfix colelem" id="u3505"><!-- column -->
+
       <p id="u4253-2">Lane 1 &amp; 17: 1kb DNA marker</p>
−
     <div class="clearfix colelem" id="u3385-9"><!-- content -->
+
       <p id="u4253-4">Lane 2&#45;8: Colony PCR with primers of Ptac (pGEX)&#45;XbaI&#45;F and VR to check Ptac&#45;GST&#45;SR/pSB1C3 (1731 bp)</p>
−
       <p id="u3385-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256004">pSB1C3&#45;Plac&#45;SS&#45;Cecropin&#45;MprF (BBa_K1256004)</a></p>
+
       <p id="u4253-6">Lane 9&#45;16: Colony PCR with primers of Pt7(pET)&#45;EcoRI&#45;XbaI&#45;F and VR to check Pt7&#45;SRPK&#45;His/pSB1C3 (2452 bp)</p>
−
       <p id="u3385-5">&nbsp;</p>
+
−
      <p id="u3385-7">MprF of Bacillus subtilis (DB2)added a RBS (BBa_B0034)was amplified by PCR and cloned into the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003) using NheI and BamHI sites. Next, the cds of cecropin was synthesized on the primers and subjeted to Gibson Assembly to make this plasmid. The amino acid sequences of cecropin was obtained from Sigma&#45;Aldrich, and the nucleotide sequence was optimized for engineering Escherichia coli using the codon optimization tool provided by Integrated DNA Technologies (IDT).</p>
+
−
    </div>
+
−
    <div class="clip_frame colelem" id="u3494"><!-- image -->
+
−
      <img class="block" id="u3494_img" src="https://static.igem.org/mediawiki/2014hs/e/ec/Psb1c3-plac-ss-cecropin-mprf.png" alt="" width="510" height="174"/>
+
−
    </div>
+
−
    </div>
+
−
    <a class="anchor_item colelem" id="bb05"></a>
+
−
    <div class="clearfix colelem" id="u3551"><!-- group -->
+
−
    <div class="clearfix grpelem" id="u3398-11"><!-- content -->
+
−
      <p id="u3398-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256005">pSB1C3&#45;Plac&#45;SS&#45;Magainin&#45;MprF (BBa_K1256005)</a></p>
+
−
       <p id="u3398-5">&nbsp;</p>
+
−
      <p id="u3398-7">MprF of Bacillus subtilis (DB2)added a RBS (BBa_B0034)was amplified by PCR and cloned into the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003) using NheI and BamHI sites. Next, the cds of magainin was synthesized on the primers and subjeted to Gibson Assembly to make this plasmid. The amino acid sequences of magainin was obtained from Sigma&#45;Aldrich, and the nucleotide sequence was optimized for engineering Escherichia coli using the codon optimization tool provided by Integrated DNA Technologies (IDT).</p>
+
−
      <p>&nbsp;</p>
+
 
       <p>&nbsp;</p>
 
       <p>&nbsp;</p>
−
    </div>
 
−
    <div class="clip_frame grpelem" id="u3499"><!-- image -->
 
−
      <img class="block" id="u3499_img" src="https://static.igem.org/mediawiki/2014hs/b/b0/Psb1c3-plac-ss-magainin-mprf.png" alt="" width="510" height="173"/>
 
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pdecomposing"><!-- group -->
+
     <div class="clearfix colelem" id="u4311-4"><!-- content -->
−
    <a class="anchor_item grpelem" id="decomposing"></a>
+
    <p>(5) Restriction enzyme check for SRPK/pSB1C3, SR/pSB1C3 and Pt7&#45;SRPK&#45;His/pSB1C3</p>
−
    <div class="clearfix grpelem" id="u3550"><!-- group -->
+
−
      <div class="clearfix grpelem" id="u3410-5"><!-- content -->
+
−
      <p>Colony PCR was performed to check the plasmid (Right Photo). The cecropin and magainin sequences were further confirmed by sequencing.</p>
+
−
      <p>&nbsp;</p>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3405"><!-- image -->
+
−
      <img class="block" id="u3405_img" src="https://static.igem.org/mediawiki/2014hs/c/c1/Gel_attack.png" alt="" width="360" height="159"/>
+
−
      </div>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pbb06"><!-- group -->
+
     <div class="clearfix colelem" id="pu4313"><!-- group -->
−
    <a class="anchor_item grpelem" id="bb06"></a>
+
     <div class="clip_frame grpelem" id="u4313"><!-- image -->
−
     <div class="clearfix grpelem" id="u3401"><!-- column -->
+
       <img class="block" id="u4313_img" src="https://static.igem.org/mediawiki/2015/f/fb/Mingdao-t1-10-crop-u4313.png" alt="" width="244" height="307"/>
−
       <div class="clearfix colelem" id="u3402-4"><!-- content -->
+
−
      <p>Decomposing</p>
+
−
      </div>
+
−
      <div class="colelem" id="u3403"><!-- simple frame --></div>
+
 
     </div>
 
     </div>
−
    </div>
+
    <div class="clip_frame grpelem" id="u4319"><!-- image -->
−
    <div class="clearfix colelem" id="u3543"><!-- column -->
+
      <img class="block" id="u4319_img" src="https://static.igem.org/mediawiki/2015/7/70/Mingdao-t1-11-crop-u4319.png" alt="" width="146" height="308"/>
−
    <div class="clearfix colelem" id="u3411-7"><!-- content -->
+
−
      <p id="u3411-2"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256006">pSB1C3&#45;Plac&#45;SS&#45;Protease (BBa_K1256006)</a></p>
+
−
      <p id="u3411-3">&nbsp;</p>
+
−
      <p id="u3411-5">The protease coding sequence (cds) of Stenotrophomonas maltophilia K279a was obtained from KEGG Genes Database. The gene was cut by NheI and BamHI and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003). The cds can be easily exchanged with NheI and BamHI. However, PstI site can no longer be used in this plasmid due to three more sites on the gene coding region.</p>
+
 
     </div>
 
     </div>
−
     <div class="clip_frame colelem" id="u3533"><!-- image -->
+
     <div class="clearfix grpelem" id="u4312-23"><!-- content -->
−
       <img class="block" id="u3533_img" src="https://static.igem.org/mediawiki/2014hs/b/b5/Psb1c3-plac-ss-protease.png" alt="" width="510" height="176"/>
+
       <p>Left panel:</p>
 +
      <p>1. SRPK/pSB1C3: EcoRI → 4036 bp</p>
 +
      <p>2. SRPK/pSB1C3: PstI → 2747 + 1238 + 51 bp</p>
 +
      <p>3. 1kb DNA marker</p>
 +
      <p>4. SR/pSB1C3: XbaI → 2815 bp</p>
 +
      <p>5. SR/pSB1C3: EcoRI + PstI → 2029 + 786 bp</p>
 +
      <p>&nbsp;</p>
 +
      <p>Right panel:</p>
 +
      <p>1. Pt7&#45;SRPK&#45;His/pSB1C3: EcoRI → 4326 bp</p>
 +
      <p>2. Pt7&#45;SRPK&#45;His/pSB1C3: PstI → 2980 + 1295 + 51 bp</p>
 +
      <p>3. 1kb DNA marker</p>
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
    <a class="anchor_item colelem" id="bb07"></a>
+
     <div class="clearfix colelem" id="u4325-4"><!-- content -->
−
     <div class="clearfix colelem" id="u3544"><!-- column -->
+
    <p>(6) Restriction enzyme check for Ptac&#45;GST&#45;SR/pSB1C3</p>
−
    <div class="clearfix colelem" id="u3412-9"><!-- content -->
+
−
      <p id="u3412-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256007">pSB1C3&#45;Plac&#45;SS&#45;Lipase (BBa_K1256007)</a></p>
+
−
      <p id="u3412-5">&nbsp;</p>
+
−
      <p id="u3412-7">The lipase coding sequence (cds) of Stenotrophomonas maltophilia K279a was obtained from KEGG Genes Database. The gene was cut by NheI and NsiI and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003) with NheI and PstI, leaving a scar of NsiI/PsiI in the end.</p>
+
−
    </div>
+
−
    <div class="clip_frame colelem" id="u3528"><!-- image -->
+
−
      <img class="block" id="u3528_img" src="https://static.igem.org/mediawiki/2014hs/d/da/Psb1c3-plac-ss-lipase.png" alt="" width="510" height="179"/>
+
−
    </div>
+
 
     </div>
 
     </div>
−
    <a class="anchor_item colelem" id="bb08"></a>
+
     <div class="clearfix colelem" id="pu4327"><!-- group -->
−
     <div class="clearfix colelem" id="u3545"><!-- column -->
+
     <div class="clip_frame grpelem" id="u4327"><!-- image -->
−
     <div class="clearfix colelem" id="u3414-9"><!-- content -->
+
       <img class="block" id="u4327_img" src="https://static.igem.org/mediawiki/2015/7/75/Mingdao-t1-12-crop-u4327.png" alt="" width="193" height="254"/>
−
       <p id="u3414-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256008">pSB1C3&#45;Plac&#45;SS&#45;DNase (BBa_K1256008)</a></p>
+
−
      <p id="u3414-5">&nbsp;</p>
+
−
      <p id="u3414-7">The DNase coding sequence (cds) of Stenotrophomonas maltophilia D457 was obtained from KEGG Genes Database. The gene was cut by NheI and BamHI and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003). The cds can be easily exchanged with NheI and BamHI. However, PstI site can no longer be used in this plasmid due to two more sites on the gene coding region.</p>
+
 
     </div>
 
     </div>
−
     <div class="clip_frame colelem" id="u3518"><!-- image -->
+
     <div class="clearfix grpelem" id="u4326-12"><!-- content -->
−
       <img class="block" id="u3518_img" src="https://static.igem.org/mediawiki/2014hs/e/ef/Psb1c3-plac-ss-dnase.png" alt="" width="510" height="176"/>
+
       <p>Lanes:</p>
 +
      <p>1. 1kb DNA marker</p>
 +
      <p>2. Ptac&#45;GST&#45;SR/pSB1C3: uncut</p>
 +
      <p>3. Ptac&#45;GST&#45;SR/pSB1C3: BamHI → 3614 bp</p>
 +
      <p>4. Ptac&#45;GST&#45;SR/pSB1C3: EcoRI + PstI → 1581 + 2033 bp</p>
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
    <a class="anchor_item colelem" id="bb09"></a>
+
     <div class="clearfix colelem" id="u4333-4"><!-- content -->
−
     <div class="clearfix colelem" id="u3546"><!-- column -->
+
    <p>(7) Sequencing data with primers of VF2 and VR</p>
−
    <div class="clearfix colelem" id="u3415-9"><!-- content -->
+
−
      <p id="u3415-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256009">pSB1C3&#45;Plac&#45;SS&#45;RNase (BBa_K1256009)</a></p>
+
−
      <p id="u3415-5">&nbsp;</p>
+
−
      <p id="u3415-7">The RNase coding sequence (cds) of Stenotrophomonas maltophilia K279a was obtained from KEGG Genes Database. The gene was cut by NheI and BglII and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003) cut by NheI and BamHI. PstI site can no longer be used in this plasmid due to one more site on the gene coding region.</p>
+
−
    </div>
+
−
    <div class="clip_frame colelem" id="u3538"><!-- image -->
+
−
      <img class="block" id="u3538_img" src="https://static.igem.org/mediawiki/2014hs/3/3f/Psb1c3-plac-ss-rnase.png" alt="" width="510" height="171"/>
+
−
    </div>
+
 
     </div>
 
     </div>
−
    <a class="anchor_item colelem" id="bb10"></a>
+
     <div class="clearfix colelem" id="u4334-16"><!-- content -->
−
     <div class="clearfix colelem" id="u3547"><!-- column -->
+
     <p id="u4334-2">a. SR/pSB1C3 (VF2 &amp; VR)</p>
−
     <div class="clearfix colelem" id="u3419-9"><!-- content -->
+
    <p id="u4334-4">b. SRPK/pSB1C3 (VF2 &amp; VR)</p>
−
      <p id="u3419-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256010">pSB1C3&#45;Plac&#45;SS&#45;Chitinase (BBa_K1256010)</a></p>
+
    <p id="u4334-6">c. Ptac&#45;GST&#45;SR/pSB1C3 (VF2 &amp; VR)</p>
−
      <p id="u3419-5">&nbsp;</p>
+
    <p id="u4334-8">d. Pt7&#45;SRPK&#45;His/pSB1C3 (VF2 &amp; VR)</p>
−
      <p id="u3419-7">The chitinase coding sequence (cds) of Stenotrophomonas maltophilia K279a was obtained from KEGG Genes Database. The gene was cut by NheI and NsiI and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003) with NheI and PstI, leaving a scar of NsiI/PsiI in the end.</p>
+
     <p id="u4334-9">&nbsp;</p>
−
     </div>
+
     <p id="u4334-13"><a class="nonblock" href="https://drive.google.com/folderview?id=0BwcRY32qizTTTDdXbENKM05KZXc&amp;usp=sharing">Download</a></p>
−
     <div class="clip_frame colelem" id="u3513"><!-- image -->
+
     <p>&nbsp;</p>
−
      <img class="block" id="u3513_img" src="https://static.igem.org/mediawiki/2014hs/4/4c/Psb1c3-plac-ss-chitinase.png" alt="" width="510" height="175"/>
+
−
     </div>
+
 
     </div>
 
     </div>
−
     <a class="anchor_item colelem" id="bb11"></a>
+
     <a class="anchor_item colelem" id="vector"></a>
−
     <div class="clearfix colelem" id="u3548"><!-- column -->
+
     <img class="colelem" id="u4335-4" alt="Vector" width="398" height="110" src="https://static.igem.org/mediawiki/2015/1/13/Mingdao-u4335-4.png"/><!-- rasterized frame -->
−
    <div class="clearfix colelem" id="u3420-9"><!-- content -->
+
    <div class="clearfix colelem" id="u4336-4"><!-- content -->
−
      <p id="u3420-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256011">pSB1C3&#45;Plac&#45;SS&#45;Glucanase (BBa_K1256011)</a></p>
+
    <p>In order to express proteins abundantly and efficiently, we transferred the genes of SR and SRPK into two commercial expression vectors of pGEX&#45;2T and pET&#45;29b, respectively.</p>
−
      <p id="u3420-5">&nbsp;</p>
+
−
      <p id="u3420-7">The glucanase coding sequence (cds) of Bacillus subtilis 168 was obtained from KEGG Genes Database. The gene was cut by NheI and BamHI and ligated to the vector of pSB1C3&#45;Plac&#45;SS&#45;NB (Part:BBa_K1256003). The cds can be easily exchanged with other BioBrick parts with NheI/PstI and SpeI/PstI, or be considered as a standard BioBrick parts (E&#45;X&#45;S&#45;P).</p>
+
−
    </div>
+
−
    <div class="clip_frame colelem" id="u3523"><!-- image -->
+
−
      <img class="block" id="u3523_img" src="https://static.igem.org/mediawiki/2014hs/a/a0/Psb1c3-plac-ss-glucanase.png" alt="" width="510" height="174"/>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pselfdestruct"><!-- group -->
+
     <a class="anchor_item colelem" id="sr-pgex-2t"></a>
−
    <a class="anchor_item grpelem" id="selfdestruct"></a>
+
    <div class="clearfix colelem" id="u4337-9"><!-- content -->
−
    <div class="clearfix grpelem" id="u3549"><!-- column -->
+
    <p id="u4337-2">SR/pGEX&#45;2T</p>
−
      <div class="clearfix colelem" id="u3422-12"><!-- content -->
+
    <p id="u4337-4">We cloned SR gene with BamHI and XbaI onto pGEX&#45;2T, in which the gene expression is fused with GST tag in the 5’ end and driven under Ptac promoter as well as regulated by lac operator. The pGEX&#45;2T backbone contains lacI repressor gene, pBR322 replication origin and ampicillin resistance cassette (AmpR).</p>
−
      <p>Genes of decomposing enzymes were cloned either from Stenotrophomonas maltophilia or Bacillus subtilis. Figure 1 showed the genes were amplified by PCR.</p>
+
    <p id="u4337-6">Plasmid map:</p>
−
      <p>&nbsp;</p>
+
    <p>&nbsp;</p>
−
      <p>Figure 1. PCR to amplify the genes of decomposing enzymes</p>
+
−
      <p>&nbsp;</p>
+
−
      <p id="u3422-10">(A<span id="u3422-8">)</span>(B)</p>
+
−
      </div>
+
−
      <div class="clearfix colelem" id="pu3423"><!-- group -->
+
−
      <div class="clip_frame grpelem" id="u3423"><!-- image -->
+
−
        <img class="block" id="u3423_img" src="https://static.igem.org/mediawiki/2014hs/b/b7/Gel_fig_b.png" alt="" width="367" height="249"/>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3428"><!-- image -->
+
−
        <img class="block" id="u3428_img" src="https://static.igem.org/mediawiki/2014hs/c/c8/Gel_fig_1.png" alt="" width="377" height="274"/>
+
−
      </div>
+
−
      </div>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pbb12"><!-- group -->
+
     <div class="clip_frame colelem" id="u4338"><!-- image -->
−
     <a class="anchor_item grpelem" id="bb12"></a>
+
     <img class="block" id="u4338_img" src="https://static.igem.org/mediawiki/2015/2/24/Mingdao-t1-13.jpeg" alt="" width="489" height="395"/>
−
    <div class="clearfix grpelem" id="u3433"><!-- column -->
+
−
      <div class="clearfix colelem" id="u3434-4"><!-- content -->
+
−
      <p>Self&#45;destruction</p>
+
−
      </div>
+
−
      <div class="colelem" id="u3435"><!-- simple frame --></div>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="u3486"><!-- column -->
+
     <a class="anchor_item colelem" id="srpk-pet-29b"></a>
−
    <div class="clearfix colelem" id="u3468"><!-- group -->
+
    <div class="clearfix colelem" id="u4344-8"><!-- content -->
−
      <div class="clearfix grpelem" id="u3446-11"><!-- content -->
+
    <p id="u4344-2">SRPK/pET&#45;29b</p>
−
      <p id="u3446-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256012">pSB1C3&#45;Plac&#45;RFP&#45;Cm&#45;LacI (BBa_K1256012)</a></p>
+
    <p id="u4344-4">We cloned SRPK gene with EcoRV and HindIII onto pET&#45;29b, in which the gene expression is fused with 6XHis tag in the 3’ end and driven under Pt7 promoter as well as regulated by lac operator. The pET&#45;29b backbone contains lacI repressor gene, ColE1 replication origin and kanamycin resistance cassette (KanR).</p>
−
      <p id="u3446-5">&nbsp;</p>
+
    <p id="u4344-6">Plasmid map:</p>
−
      <p id="u3446-7">RBS, LacI coding region and terminators (Part:BBa_Q04121) were amplified by PCR and cut by EcoRI and PstI. The PCR&#45;amplified and enzyme&#45;cut product was introduced to the novel designed plasmid backbone of pSB1C3&#45;Plac&#45;NB&#45;Cm&#45;MN ( Part:BBa_K1256002) with MfeI and NsiI. LacI gene is driven by Chloramphenicol promoter. RFP coding region can be exchanged using NheI and BamHI.</p>
+
−
      <p id="u3446-8">&nbsp;</p>
+
−
      <p id="u3446-9">&nbsp;</p>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3456"><!-- image -->
+
−
      <img class="block" id="u3456_img" src="https://static.igem.org/mediawiki/2014hs/e/ed/Psb1c3-plac-rfp-cm-laci.png" alt="" width="508" height="203"/>
+
−
      </div>
+
−
    </div>
+
−
    <div class="clearfix colelem" id="pu3451-9"><!-- group -->
+
−
      <div class="clearfix grpelem" id="u3451-9"><!-- content -->
+
−
      <p>Figure 1 showed the result of colony PCR (A) and the RFG gene expression induced by IPTG (B).</p>
+
−
      <p id="u3451-3">&nbsp;</p>
+
−
      <p id="u3451-7">(A<span id="u3451-5">)</span>(B)</p>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3476"><!-- image -->
+
−
      <img class="block" id="u3476_img" src="https://static.igem.org/mediawiki/2014hs/6/67/Colony_pcr.png" alt="" width="220" height="235"/>
+
−
      </div>
+
−
      <div class="clip_frame grpelem" id="u3481"><!-- image -->
+
−
      <img class="block" id="u3481_img" src="https://static.igem.org/mediawiki/2014hs/9/9a/Pellete.png" alt="" width="220" height="218"/>
+
−
      </div>
+
−
    </div>
+
 
     </div>
 
     </div>
−
     <a class="anchor_item colelem" id="bb13"></a>
+
    <div class="clip_frame colelem" id="u4345"><!-- image -->
−
     <div class="clearfix colelem" id="u3469"><!-- column -->
+
    <img class="block" id="u4345_img" src="https://static.igem.org/mediawiki/2015/7/7b/Mingdao-t1-14.jpeg" alt="" width="498" height="395"/>
−
     <div class="clearfix colelem" id="u3450-9"><!-- content -->
+
    </div>
−
       <p id="u3450-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256013">pSB1C3&#45;Plac&#45;ccdB&#45;Cm&#45;LacI (BBa_K1256013)</a></p>
+
     <a class="anchor_item colelem" id="data2"></a>
−
       <p id="u3450-5">&nbsp;</p>
+
     <div class="clearfix colelem" id="pu4351-5"><!-- group -->
−
      <p id="u3450-7">ccdB coding region was amplified by PCR from BBa_K145151 and exchanged with RFP on the plasmid of BBa_K1256012.</p>
+
     <div class="clearfix grpelem" id="u4351-5"><!-- content -->
 +
       <p id="u4351-2">DATA</p>
 +
       <p>&nbsp;</p>
 
     </div>
 
     </div>
−
     <div class="clip_frame colelem" id="u3461"><!-- image -->
+
     <div class="clearfix grpelem" id="u4352-6"><!-- content -->
−
       <img class="block" id="u3461_img" src="https://static.igem.org/mediawiki/2014hs/f/f3/Psb1c3-plac-ccdb-cm-laci.png" alt="" width="508" height="202"/>
+
       <p id="u4352-2">Protein Induction</p>
 +
      <p id="u4352-4">We co&#45;transformed SR/pGEX&#45;2T and SRPK/pET&#45;29b into E. coli BL21, which expresses T7 RNA polymerase under LacI repressor regulation. The bacteria were cultured in LB media supplemented with ampicillin and kanamycin. IPTG was added when the bacteria grew to OD600 value at 0.6&#45;1.0. And the protein lysates were collected after 4 hours of IPTG induction. SDS&#45;PAGE, Coomassie Blue Staining and Western Blotting were performed through the manufacturers’ instruction.</p>
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
     <a class="anchor_item colelem" id="bb14"></a>
+
     <div class="clearfix colelem" id="u4353-4"><!-- content -->
−
     <div class="clearfix colelem" id="u3475"><!-- column -->
+
    <p>Coomassie Blue Staining</p>
−
     <div class="clearfix colelem" id="u3455-9"><!-- content -->
+
    </div>
−
       <p id="u3455-4"><a class="nonblock" href="http://parts.igem.org/Part:BBa_K1256014">pSB1C3&#45;Pred&#45;ccdB&#45;light sensor (BBa_K1256014)</a></p>
+
     <div class="clearfix colelem" id="pu4354"><!-- group -->
−
      <p id="u3455-5">&nbsp;</p>
+
     <div class="clip_frame grpelem" id="u4354"><!-- image -->
−
      <p>Gene expression of ccdB (BBa_K145151), a lethal gene, is regulated by the light. The light&#45;regulated system is composed of light sensor (BBa_K1017101) and light&#45;regulated promoter (BBa_K1017301) designed and constructed by NCTU&#45;Formosa in 2013. Light sensor was first amplified by PCR, cut by EcoRI and PstI, and ligated to pSB1C3&#45;Plac&#45;NB&#45;Cm&#45;MN cut by MfeI and NsiI. Then, light&#45;regulated promoter was amplified by PCR, cut by EcoRI and NheI, and ligated to the resulting vector cut by EcoRI and NheI, which maintains the standard BioBrick assembly rule as well as the flexibility of exchanging RFP cds with NheI and BamHI. The ccdB was amplified by PCR using the forward primer with RBS and assembled to the vector with NheI and BamHI.</p>
+
       <img class="block" id="u4354_img" src="https://static.igem.org/mediawiki/2015/8/8c/Mingdao-t1-15.jpeg" alt="" width="266" height="364"/>
 
     </div>
 
     </div>
−
     <div class="clip_frame colelem" id="u3470"><!-- image -->
+
     <div class="clearfix grpelem" id="u4360-16"><!-- content -->
−
       <img class="block" id="u3470_img" src="https://static.igem.org/mediawiki/2014hs/d/d1/Psb1c3-pred-ccdb-light_sensor_%28bba_k1256014%29.png" alt="" width="509" height="207"/>
+
       <p id="u4360-2">Lanes of lysates:</p>
 +
      <p id="u4360-4">1. Control: pGEX&#45;2T &amp; pET&#45;29b</p>
 +
      <p id="u4360-6">2. SR: SR/pGEX&#45;2T &amp; pET&#45;29b</p>
 +
      <p id="u4360-8">3. SRPK: pGEX&#45;2T &amp; SRPK1/pET&#45;29b</p>
 +
      <p id="u4360-9">&nbsp;</p>
 +
      <p id="u4360-11">Result:</p>
 +
      <p id="u4360-13">SR was expressed at the expected size of 53 kDa. and SRPK was expressed at the expected size of 93 kDa. In addition, free forms of GST were observed at 27 kDa in the lanes 1 &amp; 3.</p>
 +
      <p>&nbsp;</p>
 
     </div>
 
     </div>
 
     </div>
 
     </div>
−
     <div class="clearfix colelem" id="pu3440-18"><!-- group -->
+
     <div class="clearfix colelem" id="u4362-4"><!-- content -->
−
     <div class="clearfix" id="u3440-18"><!-- content -->
+
     <p>Western Blotting</p>
−
      <p id="u3440-4"><a class="nonblock nontext anim_swing pinned-colelem" href="bio-brick.html#biobrickdesign">Biobrick Design</a></p>
+
    </div>
−
      <p id="u3440-8"><a class="nonblock nontext anim_swing pinned-colelem" href="bio-brick.html#attack">Attack</a></p>
+
    <div class="clearfix colelem" id="u4363-4"><!-- content -->
−
       <p id="u3440-12"><a class="nonblock nontext anim_swing pinned-colelem" href="bio-brick.html#decomposing">Decomposing</a></p>
+
    <p>(1) SRPK&#45;His protein expression</p>
−
      <p id="u3440-16"><a class="nonblock nontext anim_swing pinned-colelem" href="bio-brick.html#selfdestruct">Self&#45;destruction</a></p>
+
    </div>
 +
    <div class="clearfix colelem" id="pu4366"><!-- group -->
 +
    <div class="clip_frame grpelem" id="u4366"><!-- image -->
 +
       <img class="block" id="u4366_img" src="https://static.igem.org/mediawiki/2015/2/2f/Mingdao-t1-16.png" alt="" width="325" height="365"/>
 
     </div>
 
     </div>
−
     <div class="clearfix" id="u6182"><!-- column -->
+
     <img class="grpelem" id="u4364-27" alt="Procedure:
−
      <div class="position_content" id="u6182_position_content">
+
↓ mouse α&#45;His primary Ab
−
      <a class="nonblock nontext anim_swing pinned-colelem" id="u6184" href="bio-brick.html#top"><!-- rasterized frame --><img id="u6184_img" src="https://static.igem.org/mediawiki/2014hs/2/27/To_top_icon-u6165.png" alt="" width="71" height="71"/></a>
+
↓ goat α&#45;mouse IgG&#45;AP secondary Ab
−
      <a class="nonblock nontext anim_swing clearfix pinned-colelem" id="u6183-4" href="bio-brick.html#top"><!-- content --><p>Top</p></a>
+
↓ BCIP/NBT chromogen Lanes of lysates:
−
      </div>
+
1. Control: pGEX&#45;2T &amp; pET&#45;29b
 +
2. SR: SR/pGEX&#45;2T &amp; pET&#45;29b 3. SRPK: pGEX&#45;2T &amp; SRPK1/pET&#45;29b
 +
4. P&#45;SR: SR/pGEX&#45;2T &amp; SRPK1/pET&#45;29b Result:
 +
SRPK&#45;His was detected by anti&#45;His antibody at the expected size of 93 kDa, as shown in the lanes 3 &amp; 4.
 +
" width="481" height="353" src="https://static.igem.org/mediawiki/2015/7/71/Mingdao-u4364-27.png"/><!-- rasterized frame -->
 +
    </div>
 +
    <div class="clearfix colelem" id="u4372-4"><!-- content -->
 +
    <p>(2) GST&#45;SR protein expression</p>
 +
    </div>
 +
    <div class="clearfix colelem" id="pu4374"><!-- group -->
 +
    <div class="clip_frame grpelem" id="u4374"><!-- image -->
 +
      <img class="block" id="u4374_img" src="https://static.igem.org/mediawiki/2015/2/2f/Mingdao-t1-16.png" alt="" width="325" height="365"/>
 
     </div>
 
     </div>
 +
    <img class="grpelem" id="u4373-22" alt="Procedure:
 +
↓ rabbit α&#45;GST primary Ab
 +
↓ goat α&#45;rabbit IgG&#45;HRP secondary Ab
 +
↓ DAB chromogen Lanes of lysates:
 +
1. Control: pGEX&#45;2T &amp; pET&#45;29b
 +
2. SR: SR/pGEX&#45;2T &amp; pET&#45;29b 3. SRPK: pGEX&#45;2T &amp; SRPK1/pET&#45;29b
 +
4. P&#45;SR: SR/pGEX&#45;2T &amp; SRPK1/pET&#45;29b
 +
" width="481" height="257" src="https://static.igem.org/mediawiki/2015/4/4d/Mingdao-u4373-22.png"/><!-- rasterized frame -->
 +
    </div>
 +
    <div class="clearfix colelem" id="u4377-12"><!-- content -->
 +
    <p id="u4377-2">Result</p>
 +
    <p id="u4377-4">GST&#45;SR was detected by anti&#45;GST antibody at the expected size of 53 kDa, as shown in the lanes 2 &amp; 4. It is worth to note that the bands in lane 4 were shifted around 53 kDa, indicating protein modification of GST&#45;SR possibly by SRPK kinase phosphorylation (SR and SRPK exist in the bacteria all together). Moreover, the free forms of GST around 27 kDa were also observed.</p>
 +
    <p id="u4377-5">&nbsp;</p>
 +
    <p id="u4377-7">Summary</p>
 +
    <p id="u4377-9">We cloned the genes encoding SR and SRPK to BioBricks (pSB1C3) and expression vectors (pGEX&#45;2T and pET&#45;29b). After protein induction by IPTG in E. coli BL21, the SR and SRPK as well as phosphorylated SR proteins were detected in SDS&#45;PAGE with Coomassie Blue staining and Western Blotting by anti&#45;His antibody and anti&#45;GST antibody. The results showed that we successfully expressed phosphorylated SR proteins in E. coli bacteria by transformed with SR/pGEX&#45;2T and SRPK/pET&#45;29b.</p>
 +
    <p>&nbsp;</p>
 
     </div>
 
     </div>
 
     <div class="verticalspacer"></div>
 
     <div class="verticalspacer"></div>
−
    <div class="clearfix colelem" id="pu2040-3"><!-- group -->
 
−
    <div class="clearfix browser_width grpelem" id="u2040-3"><!-- content -->
 
−
      <p>&nbsp;</p>
 
−
    </div>
 
−
    <a class="nonblock nontext clip_frame grpelem" id="u1692" href="http://www.bertec.com.tw/"><!-- image --><img class="block" id="u1692_img" src="https://static.igem.org/mediawiki/2014hs/b/bf/Bertec_logo-u1692.png" alt="" width="94" height="63"/></a>
 
−
    <div class="clearfix grpelem" id="u1698-4"><!-- content -->
 
−
      <p>Sponsored by:</p>
 
−
    </div>
 
−
    <div class="clearfix grpelem" id="u1702-6"><!-- content -->
 
−
      <p><a class="nonblock" href="team.html#sponsors">and more</a></p>
 
−
    </div>
 
−
    <div class="browser_width grpelem" id="u3436"><!-- simple frame --></div>
 
−
    <div class="clearfix grpelem" id="u6138-4"><!-- content -->
 
−
      <p>Website best viewed with a 1920px wide screen with chrome</p>
 
−
    </div>
 
−
    </div>
 
 
   </div>
 
   </div>
 
   </div>
 
   </div>
 +
</div>
 
   <!-- JS includes -->
 
   <!-- JS includes -->
−
  <script type="text/javascript">
+
<script type="text/javascript">
 
   if (document.location.protocol != 'https:') document.write('\x3Cscript src="http://musecdn.businesscatalyst.com/scripts/4.0/jquery-1.8.3.min.js" type="text/javascript">\x3C/script>');
 
   if (document.location.protocol != 'https:') document.write('\x3Cscript src="http://musecdn.businesscatalyst.com/scripts/4.0/jquery-1.8.3.min.js" type="text/javascript">\x3C/script>');
 
</script>
 
</script>
−
  <script type="text/javascript">
+
<script>
−
  window.jQuery || document.write('\x3Cscript src="scripts/jquery-1.8.3.min.js" type="text/javascript">\x3C/script>');
+
/*! jQuery v1.8.3 jquery.com | jquery.org/license */
 +
(function(e,t){function _(e){var t=M[e]={};return v.each(e.split(y),function(e,n){t[n]=!0}),t}function H(e,n,r){if(r===t&&e.nodeType===1){var i="data-"+n.replace(P,"-$1").toLowerCase();r=e.getAttribute(i);if(typeof r=="string"){try{r=r==="true"?!0:r==="false"?!1:r==="null"?null:+r+""===r?+r:D.test(r)?v.parseJSON(r):r}catch(s){}v.data(e,n,r)}else r=t}return r}function B(e){var t;for(t in e){if(t==="data"&&v.isEmptyObject(e[t]))continue;if(t!=="toJSON")return!1}return!0}function et(){return!1}function tt(){return!0}function ut(e){return!e||!e.parentNode||e.parentNode.nodeType===11}function at(e,t){do e=e[t];while(e&&e.nodeType!==1);return e}function ft(e,t,n){t=t||0;if(v.isFunction(t))return v.grep(e,function(e,r){var i=!!t.call(e,r,e);return i===n});if(t.nodeType)return v.grep(e,function(e,r){return e===t===n});if(typeof t=="string"){var r=v.grep(e,function(e){return e.nodeType===1});if(it.test(t))return v.filter(t,r,!n);t=v.filter(t,r)}return v.grep(e,function(e,r){return v.inArray(e,t)>=0===n})}function lt(e){var t=ct.split("|"),n=e.createDocumentFragment();if(n.createElement)while(t.length)n.createElement(t.pop());return n}function Lt(e,t){return e.getElementsByTagName(t)[0]||e.appendChild(e.ownerDocument.createElement(t))}function At(e,t){if(t.nodeType!==1||!v.hasData(e))return;var n,r,i,s=v._data(e),o=v._data(t,s),u=s.events;if(u){delete o.handle,o.events={};for(n in u)for(r=0,i=u[n].length;r<i;r++)v.event.add(t,n,u[n][r])}o.data&&(o.data=v.extend({},o.data))}function Ot(e,t){var n;if(t.nodeType!==1)return;t.clearAttributes&&t.clearAttributes(),t.mergeAttributes&&t.mergeAttributes(e),n=t.nodeName.toLowerCase(),n==="object"?(t.parentNode&&(t.outerHTML=e.outerHTML),v.support.html5Clone&&e.innerHTML&&!v.trim(t.innerHTML)&&(t.innerHTML=e.innerHTML)):n==="input"&&Et.test(e.type)?(t.defaultChecked=t.checked=e.checked,t.value!==e.value&&(t.value=e.value)):n==="option"?t.selected=e.defaultSelected:n==="input"||n==="textarea"?t.defaultValue=e.defaultValue:n==="script"&&t.text!==e.text&&(t.text=e.text),t.removeAttribute(v.expando)}function Mt(e){return typeof e.getElementsByTagName!="undefined"?e.getElementsByTagName("*"):typeof e.querySelectorAll!="undefined"?e.querySelectorAll("*"):[]}function _t(e){Et.test(e.type)&&(e.defaultChecked=e.checked)}function Qt(e,t){if(t in e)return t;var n=t.charAt(0).toUpperCase()+t.slice(1),r=t,i=Jt.length;while(i--){t=Jt[i]+n;if(t in e)return t}return r}function Gt(e,t){return e=t||e,v.css(e,"display")==="none"||!v.contains(e.ownerDocument,e)}function Yt(e,t){var n,r,i=[],s=0,o=e.length;for(;s<o;s++){n=e[s];if(!n.style)continue;i[s]=v._data(n,"olddisplay"),t?(!i[s]&&n.style.display==="none"&&(n.style.display=""),n.style.display===""&&Gt(n)&&(i[s]=v._data(n,"olddisplay",nn(n.nodeName)))):(r=Dt(n,"display"),!i[s]&&r!=="none"&&v._data(n,"olddisplay",r))}for(s=0;s<o;s++){n=e[s];if(!n.style)continue;if(!t||n.style.display==="none"||n.style.display==="")n.style.display=t?i[s]||"":"none"}return e}function Zt(e,t,n){var r=Rt.exec(t);return r?Math.max(0,r[1]-(n||0))+(r[2]||"px"):t}function en(e,t,n,r){var i=n===(r?"border":"content")?4:t==="width"?1:0,s=0;for(;i<4;i+=2)n==="margin"&&(s+=v.css(e,n+$t[i],!0)),r?(n==="content"&&(s-=parseFloat(Dt(e,"padding"+$t[i]))||0),n!=="margin"&&(s-=parseFloat(Dt(e,"border"+$t[i]+"Width"))||0)):(s+=parseFloat(Dt(e,"padding"+$t[i]))||0,n!=="padding"&&(s+=parseFloat(Dt(e,"border"+$t[i]+"Width"))||0));return s}function tn(e,t,n){var r=t==="width"?e.offsetWidth:e.offsetHeight,i=!0,s=v.support.boxSizing&&v.css(e,"boxSizing")==="border-box";if(r<=0||r==null){r=Dt(e,t);if(r<0||r==null)r=e.style[t];if(Ut.test(r))return r;i=s&&(v.support.boxSizingReliable||r===e.style[t]),r=parseFloat(r)||0}return r+en(e,t,n||(s?"border":"content"),i)+"px"}function nn(e){if(Wt[e])return Wt[e];var t=v("<"+e+">").appendTo(i.body),n=t.css("display");t.remove();if(n==="none"||n===""){Pt=i.body.appendChild(Pt||v.extend(i.createElement("iframe"),{frameBorder:0,width:0,height:0}));if(!Ht||!Pt.createElement)Ht=(Pt.contentWindow||Pt.contentDocument).document,Ht.write("<!doctype html><html><body>"),Ht.close();t=Ht.body.appendChild(Ht.createElement(e)),n=Dt(t,"display"),i.body.removeChild(Pt)}return Wt[e]=n,n}function fn(e,t,n,r){var i;if(v.isArray(t))v.each(t,function(t,i){n||sn.test(e)?r(e,i):fn(e+"["+(typeof i=="object"?t:"")+"]",i,n,r)});else if(!n&&v.type(t)==="object")for(i in t)fn(e+"["+i+"]",t[i],n,r);else r(e,t)}function Cn(e){return function(t,n){typeof t!="string"&&(n=t,t="*");var r,i,s,o=t.toLowerCase().split(y),u=0,a=o.length;if(v.isFunction(n))for(;u<a;u++)r=o[u],s=/^\+/.test(r),s&&(r=r.substr(1)||"*"),i=e[r]=e[r]||[],i[s?"unshift":"push"](n)}}function kn(e,n,r,i,s,o){s=s||n.dataTypes[0],o=o||{},o[s]=!0;var u,a=e[s],f=0,l=a?a.length:0,c=e===Sn;for(;f<l&&(c||!u);f++)u=a[f](n,r,i),typeof u=="string"&&(!c||o[u]?u=t:(n.dataTypes.unshift(u),u=kn(e,n,r,i,u,o)));return(c||!u)&&!o["*"]&&(u=kn(e,n,r,i,"*",o)),u}function Ln(e,n){var r,i,s=v.ajaxSettings.flatOptions||{};for(r in n)n[r]!==t&&((s[r]?e:i||(i={}))[r]=n[r]);i&&v.extend(!0,e,i)}function An(e,n,r){var i,s,o,u,a=e.contents,f=e.dataTypes,l=e.responseFields;for(s in l)s in r&&(n[l[s]]=r[s]);while(f[0]==="*")f.shift(),i===t&&(i=e.mimeType||n.getResponseHeader("content-type"));if(i)for(s in a)if(a[s]&&a[s].test(i)){f.unshift(s);break}if(f[0]in r)o=f[0];else{for(s in r){if(!f[0]||e.converters[s+" "+f[0]]){o=s;break}u||(u=s)}o=o||u}if(o)return o!==f[0]&&f.unshift(o),r[o]}function On(e,t){var n,r,i,s,o=e.dataTypes.slice(),u=o[0],a={},f=0;e.dataFilter&&(t=e.dataFilter(t,e.dataType));if(o[1])for(n in e.converters)a[n.toLowerCase()]=e.converters[n];for(;i=o[++f];)if(i!=="*"){if(u!=="*"&&u!==i){n=a[u+" "+i]||a["* "+i];if(!n)for(r in a){s=r.split(" ");if(s[1]===i){n=a[u+" "+s[0]]||a["* "+s[0]];if(n){n===!0?n=a[r]:a[r]!==!0&&(i=s[0],o.splice(f--,0,i));break}}}if(n!==!0)if(n&&e["throws"])t=n(t);else try{t=n(t)}catch(l){return{state:"parsererror",error:n?l:"No conversion from "+u+" to "+i}}}u=i}return{state:"success",data:t}}function Fn(){try{return new e.XMLHttpRequest}catch(t){}}function In(){try{return new e.ActiveXObject("Microsoft.XMLHTTP")}catch(t){}}function $n(){return setTimeout(function(){qn=t},0),qn=v.now()}function Jn(e,t){v.each(t,function(t,n){var r=(Vn[t]||[]).concat(Vn["*"]),i=0,s=r.length;for(;i<s;i++)if(r[i].call(e,t,n))return})}function Kn(e,t,n){var r,i=0,s=0,o=Xn.length,u=v.Deferred().always(function(){delete a.elem}),a=function(){var t=qn||$n(),n=Math.max(0,f.startTime+f.duration-t),r=n/f.duration||0,i=1-r,s=0,o=f.tweens.length;for(;s<o;s++)f.tweens[s].run(i);return u.notifyWith(e,[f,i,n]),i<1&&o?n:(u.resolveWith(e,[f]),!1)},f=u.promise({elem:e,props:v.extend({},t),opts:v.extend(!0,{specialEasing:{}},n),originalProperties:t,originalOptions:n,startTime:qn||$n(),duration:n.duration,tweens:[],createTween:function(t,n,r){var i=v.Tween(e,f.opts,t,n,f.opts.specialEasing[t]||f.opts.easing);return f.tweens.push(i),i},stop:function(t){var n=0,r=t?f.tweens.length:0;for(;n<r;n++)f.tweens[n].run(1);return t?u.resolveWith(e,[f,t]):u.rejectWith(e,[f,t]),this}}),l=f.props;Qn(l,f.opts.specialEasing);for(;i<o;i++){r=Xn[i].call(f,e,l,f.opts);if(r)return r}return Jn(f,l),v.isFunction(f.opts.start)&&f.opts.start.call(e,f),v.fx.timer(v.extend(a,{anim:f,queue:f.opts.queue,elem:e})),f.progress(f.opts.progress).done(f.opts.done,f.opts.complete).fail(f.opts.fail).always(f.opts.always)}function Qn(e,t){var n,r,i,s,o;for(n in e){r=v.camelCase(n),i=t[r],s=e[n],v.isArray(s)&&(i=s[1],s=e[n]=s[0]),n!==r&&(e[r]=s,delete e[n]),o=v.cssHooks[r];if(o&&"expand"in o){s=o.expand(s),delete e[r];for(n in s)n in e||(e[n]=s[n],t[n]=i)}else t[r]=i}}function Gn(e,t,n){var r,i,s,o,u,a,f,l,c,h=this,p=e.style,d={},m=[],g=e.nodeType&&Gt(e);n.queue||(l=v._queueHooks(e,"fx"),l.unqueued==null&&(l.unqueued=0,c=l.empty.fire,l.empty.fire=function(){l.unqueued||c()}),l.unqueued++,h.always(function(){h.always(function(){l.unqueued--,v.queue(e,"fx").length||l.empty.fire()})})),e.nodeType===1&&("height"in t||"width"in t)&&(n.overflow=[p.overflow,p.overflowX,p.overflowY],v.css(e,"display")==="inline"&&v.css(e,"float")==="none"&&(!v.support.inlineBlockNeedsLayout||nn(e.nodeName)==="inline"?p.display="inline-block":p.zoom=1)),n.overflow&&(p.overflow="hidden",v.support.shrinkWrapBlocks||h.done(function(){p.overflow=n.overflow[0],p.overflowX=n.overflow[1],p.overflowY=n.overflow[2]}));for(r in t){s=t[r];if(Un.exec(s)){delete t[r],a=a||s==="toggle";if(s===(g?"hide":"show"))continue;m.push(r)}}o=m.length;if(o){u=v._data(e,"fxshow")||v._data(e,"fxshow",{}),"hidden"in u&&(g=u.hidden),a&&(u.hidden=!g),g?v(e).show():h.done(function(){v(e).hide()}),h.done(function(){var t;v.removeData(e,"fxshow",!0);for(t in d)v.style(e,t,d[t])});for(r=0;r<o;r++)i=m[r],f=h.createTween(i,g?u[i]:0),d[i]=u[i]||v.style(e,i),i in u||(u[i]=f.start,g&&(f.end=f.start,f.start=i==="width"||i==="height"?1:0))}}function Yn(e,t,n,r,i){return new Yn.prototype.init(e,t,n,r,i)}function Zn(e,t){var n,r={height:e},i=0;t=t?1:0;for(;i<4;i+=2-t)n=$t[i],r["margin"+n]=r["padding"+n]=e;return t&&(r.opacity=r.width=e),r}function tr(e){return v.isWindow(e)?e:e.nodeType===9?e.defaultView||e.parentWindow:!1}var n,r,i=e.document,s=e.location,o=e.navigator,u=e.jQuery,a=e.$,f=Array.prototype.push,l=Array.prototype.slice,c=Array.prototype.indexOf,h=Object.prototype.toString,p=Object.prototype.hasOwnProperty,d=String.prototype.trim,v=function(e,t){return new v.fn.init(e,t,n)},m=/[\-+]?(?:\d*\.|)\d+(?:[eE][\-+]?\d+|)/.source,g=/\S/,y=/\s+/,b=/^[\s\uFEFF\xA0]+|[\s\uFEFF\xA0]+$/g,w=/^(?:[^#<]*(<[\w\W]+>)[^>]*$|#([\w\-]*)$)/,E=/^<(\w+)\s*\/?>(?:<\/\1>|)$/,S=/^[\],:{}\s]*$/,x=/(?:^|:|,)(?:\s*\[)+/g,T=/\\(?:["\\\/bfnrt]|u[\da-fA-F]{4})/g,N=/"[^"\\\r\n]*"|true|false|null|-?(?:\d\d*\.|)\d+(?:[eE][\-+]?\d+|)/g,C=/^-ms-/,k=/-([\da-z])/gi,L=function(e,t){return(t+"").toUpperCase()},A=function(){i.addEventListener?(i.removeEventListener("DOMContentLoaded",A,!1),v.ready()):i.readyState==="complete"&&(i.detachEvent("onreadystatechange",A),v.ready())},O={};v.fn=v.prototype={constructor:v,init:function(e,n,r){var s,o,u,a;if(!e)return this;if(e.nodeType)return this.context=this[0]=e,this.length=1,this;if(typeof e=="string"){e.charAt(0)==="<"&&e.charAt(e.length-1)===">"&&e.length>=3?s=[null,e,null]:s=w.exec(e);if(s&&(s[1]||!n)){if(s[1])return n=n instanceof v?n[0]:n,a=n&&n.nodeType?n.ownerDocument||n:i,e=v.parseHTML(s[1],a,!0),E.test(s[1])&&v.isPlainObject(n)&&this.attr.call(e,n,!0),v.merge(this,e);o=i.getElementById(s[2]);if(o&&o.parentNode){if(o.id!==s[2])return r.find(e);this.length=1,this[0]=o}return this.context=i,this.selector=e,this}return!n||n.jquery?(n||r).find(e):this.constructor(n).find(e)}return v.isFunction(e)?r.ready(e):(e.selector!==t&&(this.selector=e.selector,this.context=e.context),v.makeArray(e,this))},selector:"",jquery:"1.8.3",length:0,size:function(){return this.length},toArray:function(){return l.call(this)},get:function(e){return e==null?this.toArray():e<0?this[this.length+e]:this[e]},pushStack:function(e,t,n){var r=v.merge(this.constructor(),e);return r.prevObject=this,r.context=this.context,t==="find"?r.selector=this.selector+(this.selector?" ":"")+n:t&&(r.selector=this.selector+"."+t+"("+n+")"),r},each:function(e,t){return v.each(this,e,t)},ready:function(e){return v.ready.promise().done(e),this},eq:function(e){return e=+e,e===-1?this.slice(e):this.slice(e,e+1)},first:function(){return this.eq(0)},last:function(){return this.eq(-1)},slice:function(){return this.pushStack(l.apply(this,arguments),"slice",l.call(arguments).join(","))},map:function(e){return this.pushStack(v.map(this,function(t,n){return e.call(t,n,t)}))},end:function(){return this.prevObject||this.constructor(null)},push:f,sort:[].sort,splice:[].splice},v.fn.init.prototype=v.fn,v.extend=v.fn.extend=function(){var e,n,r,i,s,o,u=arguments[0]||{},a=1,f=arguments.length,l=!1;typeof u=="boolean"&&(l=u,u=arguments[1]||{},a=2),typeof u!="object"&&!v.isFunction(u)&&(u={}),f===a&&(u=this,--a);for(;a<f;a++)if((e=arguments[a])!=null)for(n in e){r=u[n],i=e[n];if(u===i)continue;l&&i&&(v.isPlainObject(i)||(s=v.isArray(i)))?(s?(s=!1,o=r&&v.isArray(r)?r:[]):o=r&&v.isPlainObject(r)?r:{},u[n]=v.extend(l,o,i)):i!==t&&(u[n]=i)}return u},v.extend({noConflict:function(t){return e.$===v&&(e.$=a),t&&e.jQuery===v&&(e.jQuery=u),v},isReady:!1,readyWait:1,holdReady:function(e){e?v.readyWait++:v.ready(!0)},ready:function(e){if(e===!0?--v.readyWait:v.isReady)return;if(!i.body)return setTimeout(v.ready,1);v.isReady=!0;if(e!==!0&&--v.readyWait>0)return;r.resolveWith(i,[v]),v.fn.trigger&&v(i).trigger("ready").off("ready")},isFunction:function(e){return v.type(e)==="function"},isArray:Array.isArray||function(e){return v.type(e)==="array"},isWindow:function(e){return e!=null&&e==e.window},isNumeric:function(e){return!isNaN(parseFloat(e))&&isFinite(e)},type:function(e){return e==null?String(e):O[h.call(e)]||"object"},isPlainObject:function(e){if(!e||v.type(e)!=="object"||e.nodeType||v.isWindow(e))return!1;try{if(e.constructor&&!p.call(e,"constructor")&&!p.call(e.constructor.prototype,"isPrototypeOf"))return!1}catch(n){return!1}var r;for(r in e);return r===t||p.call(e,r)},isEmptyObject:function(e){var t;for(t in e)return!1;return!0},error:function(e){throw new Error(e)},parseHTML:function(e,t,n){var r;return!e||typeof e!="string"?null:(typeof t=="boolean"&&(n=t,t=0),t=t||i,(r=E.exec(e))?[t.createElement(r[1])]:(r=v.buildFragment([e],t,n?null:[]),v.merge([],(r.cacheable?v.clone(r.fragment):r.fragment).childNodes)))},parseJSON:function(t){if(!t||typeof t!="string")return null;t=v.trim(t);if(e.JSON&&e.JSON.parse)return e.JSON.parse(t);if(S.test(t.replace(T,"@").replace(N,"]").replace(x,"")))return(new Function("return "+t))();v.error("Invalid JSON: "+t)},parseXML:function(n){var r,i;if(!n||typeof n!="string")return null;try{e.DOMParser?(i=new DOMParser,r=i.parseFromString(n,"text/xml")):(r=new ActiveXObject("Microsoft.XMLDOM"),r.async="false",r.loadXML(n))}catch(s){r=t}return(!r||!r.documentElement||r.getElementsByTagName("parsererror").length)&&v.error("Invalid XML: "+n),r},noop:function(){},globalEval:function(t){t&&g.test(t)&&(e.execScript||function(t){e.eval.call(e,t)})(t)},camelCase:function(e){return e.replace(C,"ms-").replace(k,L)},nodeName:function(e,t){return e.nodeName&&e.nodeName.toLowerCase()===t.toLowerCase()},each:function(e,n,r){var i,s=0,o=e.length,u=o===t||v.isFunction(e);if(r){if(u){for(i in e)if(n.apply(e[i],r)===!1)break}else for(;s<o;)if(n.apply(e[s++],r)===!1)break}else if(u){for(i in e)if(n.call(e[i],i,e[i])===!1)break}else for(;s<o;)if(n.call(e[s],s,e[s++])===!1)break;return e},trim:d&&!d.call("\ufeff\u00a0")?function(e){return e==null?"":d.call(e)}:function(e){return e==null?"":(e+"").replace(b,"")},makeArray:function(e,t){var n,r=t||[];return e!=null&&(n=v.type(e),e.length==null||n==="string"||n==="function"||n==="regexp"||v.isWindow(e)?f.call(r,e):v.merge(r,e)),r},inArray:function(e,t,n){var r;if(t){if(c)return c.call(t,e,n);r=t.length,n=n?n<0?Math.max(0,r+n):n:0;for(;n<r;n++)if(n in t&&t[n]===e)return n}return-1},merge:function(e,n){var r=n.length,i=e.length,s=0;if(typeof r=="number")for(;s<r;s++)e[i++]=n[s];else while(n[s]!==t)e[i++]=n[s++];return e.length=i,e},grep:function(e,t,n){var r,i=[],s=0,o=e.length;n=!!n;for(;s<o;s++)r=!!t(e[s],s),n!==r&&i.push(e[s]);return i},map:function(e,n,r){var i,s,o=[],u=0,a=e.length,f=e instanceof v||a!==t&&typeof a=="number"&&(a>0&&e[0]&&e[a-1]||a===0||v.isArray(e));if(f)for(;u<a;u++)i=n(e[u],u,r),i!=null&&(o[o.length]=i);else for(s in e)i=n(e[s],s,r),i!=null&&(o[o.length]=i);return o.concat.apply([],o)},guid:1,proxy:function(e,n){var r,i,s;return typeof n=="string"&&(r=e[n],n=e,e=r),v.isFunction(e)?(i=l.call(arguments,2),s=function(){return e.apply(n,i.concat(l.call(arguments)))},s.guid=e.guid=e.guid||v.guid++,s):t},access:function(e,n,r,i,s,o,u){var a,f=r==null,l=0,c=e.length;if(r&&typeof r=="object"){for(l in r)v.access(e,n,l,r[l],1,o,i);s=1}else if(i!==t){a=u===t&&v.isFunction(i),f&&(a?(a=n,n=function(e,t,n){return a.call(v(e),n)}):(n.call(e,i),n=null));if(n)for(;l<c;l++)n(e[l],r,a?i.call(e[l],l,n(e[l],r)):i,u);s=1}return s?e:f?n.call(e):c?n(e[0],r):o},now:function(){return(new Date).getTime()}}),v.ready.promise=function(t){if(!r){r=v.Deferred();if(i.readyState==="complete")setTimeout(v.ready,1);else if(i.addEventListener)i.addEventListener("DOMContentLoaded",A,!1),e.addEventListener("load",v.ready,!1);else{i.attachEvent("onreadystatechange",A),e.attachEvent("onload",v.ready);var n=!1;try{n=e.frameElement==null&&i.documentElement}catch(s){}n&&n.doScroll&&function o(){if(!v.isReady){try{n.doScroll("left")}catch(e){return setTimeout(o,50)}v.ready()}}()}}return r.promise(t)},v.each("Boolean Number String Function Array Date RegExp Object".split(" "),function(e,t){O["[object "+t+"]"]=t.toLowerCase()}),n=v(i);var M={};v.Callbacks=function(e){e=typeof e=="string"?M[e]||_(e):v.extend({},e);var n,r,i,s,o,u,a=[],f=!e.once&&[],l=function(t){n=e.memory&&t,r=!0,u=s||0,s=0,o=a.length,i=!0;for(;a&&u<o;u++)if(a[u].apply(t[0],t[1])===!1&&e.stopOnFalse){n=!1;break}i=!1,a&&(f?f.length&&l(f.shift()):n?a=[]:c.disable())},c={add:function(){if(a){var t=a.length;(function r(t){v.each(t,function(t,n){var i=v.type(n);i==="function"?(!e.unique||!c.has(n))&&a.push(n):n&&n.length&&i!=="string"&&r(n)})})(arguments),i?o=a.length:n&&(s=t,l(n))}return this},remove:function(){return a&&v.each(arguments,function(e,t){var n;while((n=v.inArray(t,a,n))>-1)a.splice(n,1),i&&(n<=o&&o--,n<=u&&u--)}),this},has:function(e){return v.inArray(e,a)>-1},empty:function(){return a=[],this},disable:function(){return a=f=n=t,this},disabled:function(){return!a},lock:function(){return f=t,n||c.disable(),this},locked:function(){return!f},fireWith:function(e,t){return t=t||[],t=[e,t.slice?t.slice():t],a&&(!r||f)&&(i?f.push(t):l(t)),this},fire:function(){return c.fireWith(this,arguments),this},fired:function(){return!!r}};return c},v.extend({Deferred:function(e){var t=[["resolve","done",v.Callbacks("once memory"),"resolved"],["reject","fail",v.Callbacks("once memory"),"rejected"],["notify","progress",v.Callbacks("memory")]],n="pending",r={state:function(){return n},always:function(){return i.done(arguments).fail(arguments),this},then:function(){var e=arguments;return v.Deferred(function(n){v.each(t,function(t,r){var s=r[0],o=e[t];i[r[1]](v.isFunction(o)?function(){var e=o.apply(this,arguments);e&&v.isFunction(e.promise)?e.promise().done(n.resolve).fail(n.reject).progress(n.notify):n[s+"With"](this===i?n:this,[e])}:n[s])}),e=null}).promise()},promise:function(e){return e!=null?v.extend(e,r):r}},i={};return r.pipe=r.then,v.each(t,function(e,s){var o=s[2],u=s[3];r[s[1]]=o.add,u&&o.add(function(){n=u},t[e^1][2].disable,t[2][2].lock),i[s[0]]=o.fire,i[s[0]+"With"]=o.fireWith}),r.promise(i),e&&e.call(i,i),i},when:function(e){var t=0,n=l.call(arguments),r=n.length,i=r!==1||e&&v.isFunction(e.promise)?r:0,s=i===1?e:v.Deferred(),o=function(e,t,n){return function(r){t[e]=this,n[e]=arguments.length>1?l.call(arguments):r,n===u?s.notifyWith(t,n):--i||s.resolveWith(t,n)}},u,a,f;if(r>1){u=new Array(r),a=new Array(r),f=new Array(r);for(;t<r;t++)n[t]&&v.isFunction(n[t].promise)?n[t].promise().done(o(t,f,n)).fail(s.reject).progress(o(t,a,u)):--i}return i||s.resolveWith(f,n),s.promise()}}),v.support=function(){var t,n,r,s,o,u,a,f,l,c,h,p=i.createElement("div");p.setAttribute("className","t"),p.innerHTML="  <link/><table></table><a href='/a'>a</a><input type='checkbox'/>",n=p.getElementsByTagName("*"),r=p.getElementsByTagName("a")[0];if(!n||!r||!n.length)return{};s=i.createElement("select"),o=s.appendChild(i.createElement("option")),u=p.getElementsByTagName("input")[0],r.style.cssText="top:1px;float:left;opacity:.5",t={leadingWhitespace:p.firstChild.nodeType===3,tbody:!p.getElementsByTagName("tbody").length,htmlSerialize:!!p.getElementsByTagName("link").length,style:/top/.test(r.getAttribute("style")),hrefNormalized:r.getAttribute("href")==="/a",opacity:/^0.5/.test(r.style.opacity),cssFloat:!!r.style.cssFloat,checkOn:u.value==="on",optSelected:o.selected,getSetAttribute:p.className!=="t",enctype:!!i.createElement("form").enctype,html5Clone:i.createElement("nav").cloneNode(!0).outerHTML!=="<:nav></:nav>",boxModel:i.compatMode==="CSS1Compat",submitBubbles:!0,changeBubbles:!0,focusinBubbles:!1,deleteExpando:!0,noCloneEvent:!0,inlineBlockNeedsLayout:!1,shrinkWrapBlocks:!1,reliableMarginRight:!0,boxSizingReliable:!0,pixelPosition:!1},u.checked=!0,t.noCloneChecked=u.cloneNode(!0).checked,s.disabled=!0,t.optDisabled=!o.disabled;try{delete p.test}catch(d){t.deleteExpando=!1}!p.addEventListener&&p.attachEvent&&p.fireEvent&&(p.attachEvent("onclick",h=function(){t.noCloneEvent=!1}),p.cloneNode(!0).fireEvent("onclick"),p.detachEvent("onclick",h)),u=i.createElement("input"),u.value="t",u.setAttribute("type","radio"),t.radioValue=u.value==="t",u.setAttribute("checked","checked"),u.setAttribute("name","t"),p.appendChild(u),a=i.createDocumentFragment(),a.appendChild(p.lastChild),t.checkClone=a.cloneNode(!0).cloneNode(!0).lastChild.checked,t.appendChecked=u.checked,a.removeChild(u),a.appendChild(p);if(p.attachEvent)for(l in{submit:!0,change:!0,focusin:!0})f="on"+l,c=f in p,c||(p.setAttribute(f,"return;"),c=typeof p[f]=="function"),t[l+"Bubbles"]=c;return v(function(){var n,r,s,o,u="padding:0;margin:0;border:0;display:block;overflow:hidden;",a=i.getElementsByTagName("body")[0];if(!a)return;n=i.createElement("div"),n.style.cssText="visibility:hidden;border:0;width:0;height:0;position:static;top:0;margin-top:1px",a.insertBefore(n,a.firstChild),r=i.createElement("div"),n.appendChild(r),r.innerHTML="<table><tr><td></td><td>t</td></tr></table>",s=r.getElementsByTagName("td"),s[0].style.cssText="padding:0;margin:0;border:0;display:none",c=s[0].offsetHeight===0,s[0].style.display="",s[1].style.display="none",t.reliableHiddenOffsets=c&&s[0].offsetHeight===0,r.innerHTML="",r.style.cssText="box-sizing:border-box;-moz-box-sizing:border-box;-webkit-box-sizing:border-box;padding:1px;border:1px;display:block;width:4px;margin-top:1%;position:absolute;top:1%;",t.boxSizing=r.offsetWidth===4,t.doesNotIncludeMarginInBodyOffset=a.offsetTop!==1,e.getComputedStyle&&(t.pixelPosition=(e.getComputedStyle(r,null)||{}).top!=="1%",t.boxSizingReliable=(e.getComputedStyle(r,null)||{width:"4px"}).width==="4px",o=i.createElement("div"),o.style.cssText=r.style.cssText=u,o.style.marginRight=o.style.width="0",r.style.width="1px",r.appendChild(o),t.reliableMarginRight=!parseFloat((e.getComputedStyle(o,null)||{}).marginRight)),typeof r.style.zoom!="undefined"&&(r.innerHTML="",r.style.cssText=u+"width:1px;padding:1px;display:inline;zoom:1",t.inlineBlockNeedsLayout=r.offsetWidth===3,r.style.display="block",r.style.overflow="visible",r.innerHTML="<div></div>",r.firstChild.style.width="5px",t.shrinkWrapBlocks=r.offsetWidth!==3,n.style.zoom=1),a.removeChild(n),n=r=s=o=null}),a.removeChild(p),n=r=s=o=u=a=p=null,t}();var D=/(?:\{[\s\S]*\}|\[[\s\S]*\])$/,P=/([A-Z])/g;v.extend({cache:{},deletedIds:[],uuid:0,expando:"jQuery"+(v.fn.jquery+Math.random()).replace(/\D/g,""),noData:{embed:!0,object:"clsid:D27CDB6E-AE6D-11cf-96B8-444553540000",applet:!0},hasData:function(e){return e=e.nodeType?v.cache[e[v.expando]]:e[v.expando],!!e&&!B(e)},data:function(e,n,r,i){if(!v.acceptData(e))return;var s,o,u=v.expando,a=typeof n=="string",f=e.nodeType,l=f?v.cache:e,c=f?e[u]:e[u]&&u;if((!c||!l[c]||!i&&!l[c].data)&&a&&r===t)return;c||(f?e[u]=c=v.deletedIds.pop()||v.guid++:c=u),l[c]||(l[c]={},f||(l[c].toJSON=v.noop));if(typeof n=="object"||typeof n=="function")i?l[c]=v.extend(l[c],n):l[c].data=v.extend(l[c].data,n);return s=l[c],i||(s.data||(s.data={}),s=s.data),r!==t&&(s[v.camelCase(n)]=r),a?(o=s[n],o==null&&(o=s[v.camelCase(n)])):o=s,o},removeData:function(e,t,n){if(!v.acceptData(e))return;var r,i,s,o=e.nodeType,u=o?v.cache:e,a=o?e[v.expando]:v.expando;if(!u[a])return;if(t){r=n?u[a]:u[a].data;if(r){v.isArray(t)||(t in r?t=[t]:(t=v.camelCase(t),t in r?t=[t]:t=t.split(" ")));for(i=0,s=t.length;i<s;i++)delete r[t[i]];if(!(n?B:v.isEmptyObject)(r))return}}if(!n){delete u[a].data;if(!B(u[a]))return}o?v.cleanData([e],!0):v.support.deleteExpando||u!=u.window?delete u[a]:u[a]=null},_data:function(e,t,n){return v.data(e,t,n,!0)},acceptData:function(e){var t=e.nodeName&&v.noData[e.nodeName.toLowerCase()];return!t||t!==!0&&e.getAttribute("classid")===t}}),v.fn.extend({data:function(e,n){var r,i,s,o,u,a=this[0],f=0,l=null;if(e===t){if(this.length){l=v.data(a);if(a.nodeType===1&&!v._data(a,"parsedAttrs")){s=a.attributes;for(u=s.length;f<u;f++)o=s[f].name,o.indexOf("data-")||(o=v.camelCase(o.substring(5)),H(a,o,l[o]));v._data(a,"parsedAttrs",!0)}}return l}return typeof e=="object"?this.each(function(){v.data(this,e)}):(r=e.split(".",2),r[1]=r[1]?"."+r[1]:"",i=r[1]+"!",v.access(this,function(n){if(n===t)return l=this.triggerHandler("getData"+i,[r[0]]),l===t&&a&&(l=v.data(a,e),l=H(a,e,l)),l===t&&r[1]?this.data(r[0]):l;r[1]=n,this.each(function(){var t=v(this);t.triggerHandler("setData"+i,r),v.data(this,e,n),t.triggerHandler("changeData"+i,r)})},null,n,arguments.length>1,null,!1))},removeData:function(e){return this.each(function(){v.removeData(this,e)})}}),v.extend({queue:function(e,t,n){var r;if(e)return t=(t||"fx")+"queue",r=v._data(e,t),n&&(!r||v.isArray(n)?r=v._data(e,t,v.makeArray(n)):r.push(n)),r||[]},dequeue:function(e,t){t=t||"fx";var n=v.queue(e,t),r=n.length,i=n.shift(),s=v._queueHooks(e,t),o=function(){v.dequeue(e,t)};i==="inprogress"&&(i=n.shift(),r--),i&&(t==="fx"&&n.unshift("inprogress"),delete s.stop,i.call(e,o,s)),!r&&s&&s.empty.fire()},_queueHooks:function(e,t){var n=t+"queueHooks";return v._data(e,n)||v._data(e,n,{empty:v.Callbacks("once memory").add(function(){v.removeData(e,t+"queue",!0),v.removeData(e,n,!0)})})}}),v.fn.extend({queue:function(e,n){var r=2;return typeof e!="string"&&(n=e,e="fx",r--),arguments.length<r?v.queue(this[0],e):n===t?this:this.each(function(){var t=v.queue(this,e,n);v._queueHooks(this,e),e==="fx"&&t[0]!=="inprogress"&&v.dequeue(this,e)})},dequeue:function(e){return this.each(function(){v.dequeue(this,e)})},delay:function(e,t){return e=v.fx?v.fx.speeds[e]||e:e,t=t||"fx",this.queue(t,function(t,n){var r=setTimeout(t,e);n.stop=function(){clearTimeout(r)}})},clearQueue:function(e){return this.queue(e||"fx",[])},promise:function(e,n){var r,i=1,s=v.Deferred(),o=this,u=this.length,a=function(){--i||s.resolveWith(o,[o])};typeof e!="string"&&(n=e,e=t),e=e||"fx";while(u--)r=v._data(o[u],e+"queueHooks"),r&&r.empty&&(i++,r.empty.add(a));return a(),s.promise(n)}});var j,F,I,q=/[\t\r\n]/g,R=/\r/g,U=/^(?:button|input)$/i,z=/^(?:button|input|object|select|textarea)$/i,W=/^a(?:rea|)$/i,X=/^(?:autofocus|autoplay|async|checked|controls|defer|disabled|hidden|loop|multiple|open|readonly|required|scoped|selected)$/i,V=v.support.getSetAttribute;v.fn.extend({attr:function(e,t){return v.access(this,v.attr,e,t,arguments.length>1)},removeAttr:function(e){return this.each(function(){v.removeAttr(this,e)})},prop:function(e,t){return v.access(this,v.prop,e,t,arguments.length>1)},removeProp:function(e){return e=v.propFix[e]||e,this.each(function(){try{this[e]=t,delete this[e]}catch(n){}})},addClass:function(e){var t,n,r,i,s,o,u;if(v.isFunction(e))return this.each(function(t){v(this).addClass(e.call(this,t,this.className))});if(e&&typeof e=="string"){t=e.split(y);for(n=0,r=this.length;n<r;n++){i=this[n];if(i.nodeType===1)if(!i.className&&t.length===1)i.className=e;else{s=" "+i.className+" ";for(o=0,u=t.length;o<u;o++)s.indexOf(" "+t[o]+" ")<0&&(s+=t[o]+" ");i.className=v.trim(s)}}}return this},removeClass:function(e){var n,r,i,s,o,u,a;if(v.isFunction(e))return this.each(function(t){v(this).removeClass(e.call(this,t,this.className))});if(e&&typeof e=="string"||e===t){n=(e||"").split(y);for(u=0,a=this.length;u<a;u++){i=this[u];if(i.nodeType===1&&i.className){r=(" "+i.className+" ").replace(q," ");for(s=0,o=n.length;s<o;s++)while(r.indexOf(" "+n[s]+" ")>=0)r=r.replace(" "+n[s]+" "," ");i.className=e?v.trim(r):""}}}return this},toggleClass:function(e,t){var n=typeof e,r=typeof t=="boolean";return v.isFunction(e)?this.each(function(n){v(this).toggleClass(e.call(this,n,this.className,t),t)}):this.each(function(){if(n==="string"){var i,s=0,o=v(this),u=t,a=e.split(y);while(i=a[s++])u=r?u:!o.hasClass(i),o[u?"addClass":"removeClass"](i)}else if(n==="undefined"||n==="boolean")this.className&&v._data(this,"__className__",this.className),this.className=this.className||e===!1?"":v._data(this,"__className__")||""})},hasClass:function(e){var t=" "+e+" ",n=0,r=this.length;for(;n<r;n++)if(this[n].nodeType===1&&(" "+this[n].className+" ").replace(q," ").indexOf(t)>=0)return!0;return!1},val:function(e){var n,r,i,s=this[0];if(!arguments.length){if(s)return n=v.valHooks[s.type]||v.valHooks[s.nodeName.toLowerCase()],n&&"get"in n&&(r=n.get(s,"value"))!==t?r:(r=s.value,typeof r=="string"?r.replace(R,""):r==null?"":r);return}return i=v.isFunction(e),this.each(function(r){var s,o=v(this);if(this.nodeType!==1)return;i?s=e.call(this,r,o.val()):s=e,s==null?s="":typeof s=="number"?s+="":v.isArray(s)&&(s=v.map(s,function(e){return e==null?"":e+""})),n=v.valHooks[this.type]||v.valHooks[this.nodeName.toLowerCase()];if(!n||!("set"in n)||n.set(this,s,"value")===t)this.value=s})}}),v.extend({valHooks:{option:{get:function(e){var t=e.attributes.value;return!t||t.specified?e.value:e.text}},select:{get:function(e){var t,n,r=e.options,i=e.selectedIndex,s=e.type==="select-one"||i<0,o=s?null:[],u=s?i+1:r.length,a=i<0?u:s?i:0;for(;a<u;a++){n=r[a];if((n.selected||a===i)&&(v.support.optDisabled?!n.disabled:n.getAttribute("disabled")===null)&&(!n.parentNode.disabled||!v.nodeName(n.parentNode,"optgroup"))){t=v(n).val();if(s)return t;o.push(t)}}return o},set:function(e,t){var n=v.makeArray(t);return v(e).find("option").each(function(){this.selected=v.inArray(v(this).val(),n)>=0}),n.length||(e.selectedIndex=-1),n}}},attrFn:{},attr:function(e,n,r,i){var s,o,u,a=e.nodeType;if(!e||a===3||a===8||a===2)return;if(i&&v.isFunction(v.fn[n]))return v(e)[n](r);if(typeof e.getAttribute=="undefined")return v.prop(e,n,r);u=a!==1||!v.isXMLDoc(e),u&&(n=n.toLowerCase(),o=v.attrHooks[n]||(X.test(n)?F:j));if(r!==t){if(r===null){v.removeAttr(e,n);return}return o&&"set"in o&&u&&(s=o.set(e,r,n))!==t?s:(e.setAttribute(n,r+""),r)}return o&&"get"in o&&u&&(s=o.get(e,n))!==null?s:(s=e.getAttribute(n),s===null?t:s)},removeAttr:function(e,t){var n,r,i,s,o=0;if(t&&e.nodeType===1){r=t.split(y);for(;o<r.length;o++)i=r[o],i&&(n=v.propFix[i]||i,s=X.test(i),s||v.attr(e,i,""),e.removeAttribute(V?i:n),s&&n in e&&(e[n]=!1))}},attrHooks:{type:{set:function(e,t){if(U.test(e.nodeName)&&e.parentNode)v.error("type property can't be changed");else if(!v.support.radioValue&&t==="radio"&&v.nodeName(e,"input")){var n=e.value;return e.setAttribute("type",t),n&&(e.value=n),t}}},value:{get:function(e,t){return j&&v.nodeName(e,"button")?j.get(e,t):t in e?e.value:null},set:function(e,t,n){if(j&&v.nodeName(e,"button"))return j.set(e,t,n);e.value=t}}},propFix:{tabindex:"tabIndex",readonly:"readOnly","for":"htmlFor","class":"className",maxlength:"maxLength",cellspacing:"cellSpacing",cellpadding:"cellPadding",rowspan:"rowSpan",colspan:"colSpan",usemap:"useMap",frameborder:"frameBorder",contenteditable:"contentEditable"},prop:function(e,n,r){var i,s,o,u=e.nodeType;if(!e||u===3||u===8||u===2)return;return o=u!==1||!v.isXMLDoc(e),o&&(n=v.propFix[n]||n,s=v.propHooks[n]),r!==t?s&&"set"in s&&(i=s.set(e,r,n))!==t?i:e[n]=r:s&&"get"in s&&(i=s.get(e,n))!==null?i:e[n]},propHooks:{tabIndex:{get:function(e){var n=e.getAttributeNode("tabindex");return n&&n.specified?parseInt(n.value,10):z.test(e.nodeName)||W.test(e.nodeName)&&e.href?0:t}}}}),F={get:function(e,n){var r,i=v.prop(e,n);return i===!0||typeof i!="boolean"&&(r=e.getAttributeNode(n))&&r.nodeValue!==!1?n.toLowerCase():t},set:function(e,t,n){var r;return t===!1?v.removeAttr(e,n):(r=v.propFix[n]||n,r in e&&(e[r]=!0),e.setAttribute(n,n.toLowerCase())),n}},V||(I={name:!0,id:!0,coords:!0},j=v.valHooks.button={get:function(e,n){var r;return r=e.getAttributeNode(n),r&&(I[n]?r.value!=="":r.specified)?r.value:t},set:function(e,t,n){var r=e.getAttributeNode(n);return r||(r=i.createAttribute(n),e.setAttributeNode(r)),r.value=t+""}},v.each(["width","height"],function(e,t){v.attrHooks[t]=v.extend(v.attrHooks[t],{set:function(e,n){if(n==="")return e.setAttribute(t,"auto"),n}})}),v.attrHooks.contenteditable={get:j.get,set:function(e,t,n){t===""&&(t="false"),j.set(e,t,n)}}),v.support.hrefNormalized||v.each(["href","src","width","height"],function(e,n){v.attrHooks[n]=v.extend(v.attrHooks[n],{get:function(e){var r=e.getAttribute(n,2);return r===null?t:r}})}),v.support.style||(v.attrHooks.style={get:function(e){return e.style.cssText.toLowerCase()||t},set:function(e,t){return e.style.cssText=t+""}}),v.support.optSelected||(v.propHooks.selected=v.extend(v.propHooks.selected,{get:function(e){var t=e.parentNode;return t&&(t.selectedIndex,t.parentNode&&t.parentNode.selectedIndex),null}})),v.support.enctype||(v.propFix.enctype="encoding"),v.support.checkOn||v.each(["radio","checkbox"],function(){v.valHooks[this]={get:function(e){return e.getAttribute("value")===null?"on":e.value}}}),v.each(["radio","checkbox"],function(){v.valHooks[this]=v.extend(v.valHooks[this],{set:function(e,t){if(v.isArray(t))return e.checked=v.inArray(v(e).val(),t)>=0}})});var $=/^(?:textarea|input|select)$/i,J=/^([^\.]*|)(?:\.(.+)|)$/,K=/(?:^|\s)hover(\.\S+|)\b/,Q=/^key/,G=/^(?:mouse|contextmenu)|click/,Y=/^(?:focusinfocus|focusoutblur)$/,Z=function(e){return v.event.special.hover?e:e.replace(K,"mouseenter$1 mouseleave$1")};v.event={add:function(e,n,r,i,s){var o,u,a,f,l,c,h,p,d,m,g;if(e.nodeType===3||e.nodeType===8||!n||!r||!(o=v._data(e)))return;r.handler&&(d=r,r=d.handler,s=d.selector),r.guid||(r.guid=v.guid++),a=o.events,a||(o.events=a={}),u=o.handle,u||(o.handle=u=function(e){return typeof v=="undefined"||!!e&&v.event.triggered===e.type?t:v.event.dispatch.apply(u.elem,arguments)},u.elem=e),n=v.trim(Z(n)).split(" ");for(f=0;f<n.length;f++){l=J.exec(n[f])||[],c=l[1],h=(l[2]||"").split(".").sort(),g=v.event.special[c]||{},c=(s?g.delegateType:g.bindType)||c,g=v.event.special[c]||{},p=v.extend({type:c,origType:l[1],data:i,handler:r,guid:r.guid,selector:s,needsContext:s&&v.expr.match.needsContext.test(s),namespace:h.join(".")},d),m=a[c];if(!m){m=a[c]=[],m.delegateCount=0;if(!g.setup||g.setup.call(e,i,h,u)===!1)e.addEventListener?e.addEventListener(c,u,!1):e.attachEvent&&e.attachEvent("on"+c,u)}g.add&&(g.add.call(e,p),p.handler.guid||(p.handler.guid=r.guid)),s?m.splice(m.delegateCount++,0,p):m.push(p),v.event.global[c]=!0}e=null},global:{},remove:function(e,t,n,r,i){var s,o,u,a,f,l,c,h,p,d,m,g=v.hasData(e)&&v._data(e);if(!g||!(h=g.events))return;t=v.trim(Z(t||"")).split(" ");for(s=0;s<t.length;s++){o=J.exec(t[s])||[],u=a=o[1],f=o[2];if(!u){for(u in h)v.event.remove(e,u+t[s],n,r,!0);continue}p=v.event.special[u]||{},u=(r?p.delegateType:p.bindType)||u,d=h[u]||[],l=d.length,f=f?new RegExp("(^|\\.)"+f.split(".").sort().join("\\.(?:.*\\.|)")+"(\\.|$)"):null;for(c=0;c<d.length;c++)m=d[c],(i||a===m.origType)&&(!n||n.guid===m.guid)&&(!f||f.test(m.namespace))&&(!r||r===m.selector||r==="**"&&m.selector)&&(d.splice(c--,1),m.selector&&d.delegateCount--,p.remove&&p.remove.call(e,m));d.length===0&&l!==d.length&&((!p.teardown||p.teardown.call(e,f,g.handle)===!1)&&v.removeEvent(e,u,g.handle),delete h[u])}v.isEmptyObject(h)&&(delete g.handle,v.removeData(e,"events",!0))},customEvent:{getData:!0,setData:!0,changeData:!0},trigger:function(n,r,s,o){if(!s||s.nodeType!==3&&s.nodeType!==8){var u,a,f,l,c,h,p,d,m,g,y=n.type||n,b=[];if(Y.test(y+v.event.triggered))return;y.indexOf("!")>=0&&(y=y.slice(0,-1),a=!0),y.indexOf(".")>=0&&(b=y.split("."),y=b.shift(),b.sort());if((!s||v.event.customEvent[y])&&!v.event.global[y])return;n=typeof n=="object"?n[v.expando]?n:new v.Event(y,n):new v.Event(y),n.type=y,n.isTrigger=!0,n.exclusive=a,n.namespace=b.join("."),n.namespace_re=n.namespace?new RegExp("(^|\\.)"+b.join("\\.(?:.*\\.|)")+"(\\.|$)"):null,h=y.indexOf(":")<0?"on"+y:"";if(!s){u=v.cache;for(f in u)u[f].events&&u[f].events[y]&&v.event.trigger(n,r,u[f].handle.elem,!0);return}n.result=t,n.target||(n.target=s),r=r!=null?v.makeArray(r):[],r.unshift(n),p=v.event.special[y]||{};if(p.trigger&&p.trigger.apply(s,r)===!1)return;m=[[s,p.bindType||y]];if(!o&&!p.noBubble&&!v.isWindow(s)){g=p.delegateType||y,l=Y.test(g+y)?s:s.parentNode;for(c=s;l;l=l.parentNode)m.push([l,g]),c=l;c===(s.ownerDocument||i)&&m.push([c.defaultView||c.parentWindow||e,g])}for(f=0;f<m.length&&!n.isPropagationStopped();f++)l=m[f][0],n.type=m[f][1],d=(v._data(l,"events")||{})[n.type]&&v._data(l,"handle"),d&&d.apply(l,r),d=h&&l[h],d&&v.acceptData(l)&&d.apply&&d.apply(l,r)===!1&&n.preventDefault();return n.type=y,!o&&!n.isDefaultPrevented()&&(!p._default||p._default.apply(s.ownerDocument,r)===!1)&&(y!=="click"||!v.nodeName(s,"a"))&&v.acceptData(s)&&h&&s[y]&&(y!=="focus"&&y!=="blur"||n.target.offsetWidth!==0)&&!v.isWindow(s)&&(c=s[h],c&&(s[h]=null),v.event.triggered=y,s[y](),v.event.triggered=t,c&&(s[h]=c)),n.result}return},dispatch:function(n){n=v.event.fix(n||e.event);var r,i,s,o,u,a,f,c,h,p,d=(v._data(this,"events")||{})[n.type]||[],m=d.delegateCount,g=l.call(arguments),y=!n.exclusive&&!n.namespace,b=v.event.special[n.type]||{},w=[];g[0]=n,n.delegateTarget=this;if(b.preDispatch&&b.preDispatch.call(this,n)===!1)return;if(m&&(!n.button||n.type!=="click"))for(s=n.target;s!=this;s=s.parentNode||this)if(s.disabled!==!0||n.type!=="click"){u={},f=[];for(r=0;r<m;r++)c=d[r],h=c.selector,u[h]===t&&(u[h]=c.needsContext?v(h,this).index(s)>=0:v.find(h,this,null,[s]).length),u[h]&&f.push(c);f.length&&w.push({elem:s,matches:f})}d.length>m&&w.push({elem:this,matches:d.slice(m)});for(r=0;r<w.length&&!n.isPropagationStopped();r++){a=w[r],n.currentTarget=a.elem;for(i=0;i<a.matches.length&&!n.isImmediatePropagationStopped();i++){c=a.matches[i];if(y||!n.namespace&&!c.namespace||n.namespace_re&&n.namespace_re.test(c.namespace))n.data=c.data,n.handleObj=c,o=((v.event.special[c.origType]||{}).handle||c.handler).apply(a.elem,g),o!==t&&(n.result=o,o===!1&&(n.preventDefault(),n.stopPropagation()))}}return b.postDispatch&&b.postDispatch.call(this,n),n.result},props:"attrChange attrName relatedNode srcElement altKey bubbles cancelable ctrlKey currentTarget eventPhase metaKey relatedTarget shiftKey target timeStamp view which".split(" "),fixHooks:{},keyHooks:{props:"char charCode key keyCode".split(" "),filter:function(e,t){return e.which==null&&(e.which=t.charCode!=null?t.charCode:t.keyCode),e}},mouseHooks:{props:"button buttons clientX clientY fromElement offsetX offsetY pageX pageY screenX screenY toElement".split(" "),filter:function(e,n){var r,s,o,u=n.button,a=n.fromElement;return e.pageX==null&&n.clientX!=null&&(r=e.target.ownerDocument||i,s=r.documentElement,o=r.body,e.pageX=n.clientX+(s&&s.scrollLeft||o&&o.scrollLeft||0)-(s&&s.clientLeft||o&&o.clientLeft||0),e.pageY=n.clientY+(s&&s.scrollTop||o&&o.scrollTop||0)-(s&&s.clientTop||o&&o.clientTop||0)),!e.relatedTarget&&a&&(e.relatedTarget=a===e.target?n.toElement:a),!e.which&&u!==t&&(e.which=u&1?1:u&2?3:u&4?2:0),e}},fix:function(e){if(e[v.expando])return e;var t,n,r=e,s=v.event.fixHooks[e.type]||{},o=s.props?this.props.concat(s.props):this.props;e=v.Event(r);for(t=o.length;t;)n=o[--t],e[n]=r[n];return e.target||(e.target=r.srcElement||i),e.target.nodeType===3&&(e.target=e.target.parentNode),e.metaKey=!!e.metaKey,s.filter?s.filter(e,r):e},special:{load:{noBubble:!0},focus:{delegateType:"focusin"},blur:{delegateType:"focusout"},beforeunload:{setup:function(e,t,n){v.isWindow(this)&&(this.onbeforeunload=n)},teardown:function(e,t){this.onbeforeunload===t&&(this.onbeforeunload=null)}}},simulate:function(e,t,n,r){var i=v.extend(new v.Event,n,{type:e,isSimulated:!0,originalEvent:{}});r?v.event.trigger(i,null,t):v.event.dispatch.call(t,i),i.isDefaultPrevented()&&n.preventDefault()}},v.event.handle=v.event.dispatch,v.removeEvent=i.removeEventListener?function(e,t,n){e.removeEventListener&&e.removeEventListener(t,n,!1)}:function(e,t,n){var r="on"+t;e.detachEvent&&(typeof e[r]=="undefined"&&(e[r]=null),e.detachEvent(r,n))},v.Event=function(e,t){if(!(this instanceof v.Event))return new v.Event(e,t);e&&e.type?(this.originalEvent=e,this.type=e.type,this.isDefaultPrevented=e.defaultPrevented||e.returnValue===!1||e.getPreventDefault&&e.getPreventDefault()?tt:et):this.type=e,t&&v.extend(this,t),this.timeStamp=e&&e.timeStamp||v.now(),this[v.expando]=!0},v.Event.prototype={preventDefault:function(){this.isDefaultPrevented=tt;var e=this.originalEvent;if(!e)return;e.preventDefault?e.preventDefault():e.returnValue=!1},stopPropagation:function(){this.isPropagationStopped=tt;var e=this.originalEvent;if(!e)return;e.stopPropagation&&e.stopPropagation(),e.cancelBubble=!0},stopImmediatePropagation:function(){this.isImmediatePropagationStopped=tt,this.stopPropagation()},isDefaultPrevented:et,isPropagationStopped:et,isImmediatePropagationStopped:et},v.each({mouseenter:"mouseover",mouseleave:"mouseout"},function(e,t){v.event.special[e]={delegateType:t,bindType:t,handle:function(e){var n,r=this,i=e.relatedTarget,s=e.handleObj,o=s.selector;if(!i||i!==r&&!v.contains(r,i))e.type=s.origType,n=s.handler.apply(this,arguments),e.type=t;return n}}}),v.support.submitBubbles||(v.event.special.submit={setup:function(){if(v.nodeName(this,"form"))return!1;v.event.add(this,"click._submit keypress._submit",function(e){var n=e.target,r=v.nodeName(n,"input")||v.nodeName(n,"button")?n.form:t;r&&!v._data(r,"_submit_attached")&&(v.event.add(r,"submit._submit",function(e){e._submit_bubble=!0}),v._data(r,"_submit_attached",!0))})},postDispatch:function(e){e._submit_bubble&&(delete e._submit_bubble,this.parentNode&&!e.isTrigger&&v.event.simulate("submit",this.parentNode,e,!0))},teardown:function(){if(v.nodeName(this,"form"))return!1;v.event.remove(this,"._submit")}}),v.support.changeBubbles||(v.event.special.change={setup:function(){if($.test(this.nodeName)){if(this.type==="checkbox"||this.type==="radio")v.event.add(this,"propertychange._change",function(e){e.originalEvent.propertyName==="checked"&&(this._just_changed=!0)}),v.event.add(this,"click._change",function(e){this._just_changed&&!e.isTrigger&&(this._just_changed=!1),v.event.simulate("change",this,e,!0)});return!1}v.event.add(this,"beforeactivate._change",function(e){var t=e.target;$.test(t.nodeName)&&!v._data(t,"_change_attached")&&(v.event.add(t,"change._change",function(e){this.parentNode&&!e.isSimulated&&!e.isTrigger&&v.event.simulate("change",this.parentNode,e,!0)}),v._data(t,"_change_attached",!0))})},handle:function(e){var t=e.target;if(this!==t||e.isSimulated||e.isTrigger||t.type!=="radio"&&t.type!=="checkbox")return e.handleObj.handler.apply(this,arguments)},teardown:function(){return v.event.remove(this,"._change"),!$.test(this.nodeName)}}),v.support.focusinBubbles||v.each({focus:"focusin",blur:"focusout"},function(e,t){var n=0,r=function(e){v.event.simulate(t,e.target,v.event.fix(e),!0)};v.event.special[t]={setup:function(){n++===0&&i.addEventListener(e,r,!0)},teardown:function(){--n===0&&i.removeEventListener(e,r,!0)}}}),v.fn.extend({on:function(e,n,r,i,s){var o,u;if(typeof e=="object"){typeof n!="string"&&(r=r||n,n=t);for(u in e)this.on(u,n,r,e[u],s);return this}r==null&&i==null?(i=n,r=n=t):i==null&&(typeof n=="string"?(i=r,r=t):(i=r,r=n,n=t));if(i===!1)i=et;else if(!i)return this;return s===1&&(o=i,i=function(e){return v().off(e),o.apply(this,arguments)},i.guid=o.guid||(o.guid=v.guid++)),this.each(function(){v.event.add(this,e,i,r,n)})},one:function(e,t,n,r){return this.on(e,t,n,r,1)},off:function(e,n,r){var i,s;if(e&&e.preventDefault&&e.handleObj)return i=e.handleObj,v(e.delegateTarget).off(i.namespace?i.origType+"."+i.namespace:i.origType,i.selector,i.handler),this;if(typeof e=="object"){for(s in e)this.off(s,n,e[s]);return this}if(n===!1||typeof n=="function")r=n,n=t;return r===!1&&(r=et),this.each(function(){v.event.remove(this,e,r,n)})},bind:function(e,t,n){return this.on(e,null,t,n)},unbind:function(e,t){return this.off(e,null,t)},live:function(e,t,n){return v(this.context).on(e,this.selector,t,n),this},die:function(e,t){return v(this.context).off(e,this.selector||"**",t),this},delegate:function(e,t,n,r){return this.on(t,e,n,r)},undelegate:function(e,t,n){return arguments.length===1?this.off(e,"**"):this.off(t,e||"**",n)},trigger:function(e,t){return this.each(function(){v.event.trigger(e,t,this)})},triggerHandler:function(e,t){if(this[0])return v.event.trigger(e,t,this[0],!0)},toggle:function(e){var t=arguments,n=e.guid||v.guid++,r=0,i=function(n){var i=(v._data(this,"lastToggle"+e.guid)||0)%r;return v._data(this,"lastToggle"+e.guid,i+1),n.preventDefault(),t[i].apply(this,arguments)||!1};i.guid=n;while(r<t.length)t[r++].guid=n;return this.click(i)},hover:function(e,t){return this.mouseenter(e).mouseleave(t||e)}}),v.each("blur focus focusin focusout load resize scroll unload click dblclick mousedown mouseup mousemove mouseover mouseout mouseenter mouseleave change select submit keydown keypress keyup error contextmenu".split(" "),function(e,t){v.fn[t]=function(e,n){return n==null&&(n=e,e=null),arguments.length>0?this.on(t,null,e,n):this.trigger(t)},Q.test(t)&&(v.event.fixHooks[t]=v.event.keyHooks),G.test(t)&&(v.event.fixHooks[t]=v.event.mouseHooks)}),function(e,t){function nt(e,t,n,r){n=n||[],t=t||g;var i,s,a,f,l=t.nodeType;if(!e||typeof e!="string")return n;if(l!==1&&l!==9)return[];a=o(t);if(!a&&!r)if(i=R.exec(e))if(f=i[1]){if(l===9){s=t.getElementById(f);if(!s||!s.parentNode)return n;if(s.id===f)return n.push(s),n}else if(t.ownerDocument&&(s=t.ownerDocument.getElementById(f))&&u(t,s)&&s.id===f)return n.push(s),n}else{if(i[2])return S.apply(n,x.call(t.getElementsByTagName(e),0)),n;if((f=i[3])&&Z&&t.getElementsByClassName)return S.apply(n,x.call(t.getElementsByClassName(f),0)),n}return vt(e.replace(j,"$1"),t,n,r,a)}function rt(e){return function(t){var n=t.nodeName.toLowerCase();return n==="input"&&t.type===e}}function it(e){return function(t){var n=t.nodeName.toLowerCase();return(n==="input"||n==="button")&&t.type===e}}function st(e){return N(function(t){return t=+t,N(function(n,r){var i,s=e([],n.length,t),o=s.length;while(o--)n[i=s[o]]&&(n[i]=!(r[i]=n[i]))})})}function ot(e,t,n){if(e===t)return n;var r=e.nextSibling;while(r){if(r===t)return-1;r=r.nextSibling}return 1}function ut(e,t){var n,r,s,o,u,a,f,l=L[d][e+" "];if(l)return t?0:l.slice(0);u=e,a=[],f=i.preFilter;while(u){if(!n||(r=F.exec(u)))r&&(u=u.slice(r[0].length)||u),a.push(s=[]);n=!1;if(r=I.exec(u))s.push(n=new m(r.shift())),u=u.slice(n.length),n.type=r[0].replace(j," ");for(o in i.filter)(r=J[o].exec(u))&&(!f[o]||(r=f[o](r)))&&(s.push(n=new m(r.shift())),u=u.slice(n.length),n.type=o,n.matches=r);if(!n)break}return t?u.length:u?nt.error(e):L(e,a).slice(0)}function at(e,t,r){var i=t.dir,s=r&&t.dir==="parentNode",o=w++;return t.first?function(t,n,r){while(t=t[i])if(s||t.nodeType===1)return e(t,n,r)}:function(t,r,u){if(!u){var a,f=b+" "+o+" ",l=f+n;while(t=t[i])if(s||t.nodeType===1){if((a=t[d])===l)return t.sizset;if(typeof a=="string"&&a.indexOf(f)===0){if(t.sizset)return t}else{t[d]=l;if(e(t,r,u))return t.sizset=!0,t;t.sizset=!1}}}else while(t=t[i])if(s||t.nodeType===1)if(e(t,r,u))return t}}function ft(e){return e.length>1?function(t,n,r){var i=e.length;while(i--)if(!e[i](t,n,r))return!1;return!0}:e[0]}function lt(e,t,n,r,i){var s,o=[],u=0,a=e.length,f=t!=null;for(;u<a;u++)if(s=e[u])if(!n||n(s,r,i))o.push(s),f&&t.push(u);return o}function ct(e,t,n,r,i,s){return r&&!r[d]&&(r=ct(r)),i&&!i[d]&&(i=ct(i,s)),N(function(s,o,u,a){var f,l,c,h=[],p=[],d=o.length,v=s||dt(t||"*",u.nodeType?[u]:u,[]),m=e&&(s||!t)?lt(v,h,e,u,a):v,g=n?i||(s?e:d||r)?[]:o:m;n&&n(m,g,u,a);if(r){f=lt(g,p),r(f,[],u,a),l=f.length;while(l--)if(c=f[l])g[p[l]]=!(m[p[l]]=c)}if(s){if(i||e){if(i){f=[],l=g.length;while(l--)(c=g[l])&&f.push(m[l]=c);i(null,g=[],f,a)}l=g.length;while(l--)(c=g[l])&&(f=i?T.call(s,c):h[l])>-1&&(s[f]=!(o[f]=c))}}else g=lt(g===o?g.splice(d,g.length):g),i?i(null,o,g,a):S.apply(o,g)})}function ht(e){var t,n,r,s=e.length,o=i.relative[e[0].type],u=o||i.relative[" "],a=o?1:0,f=at(function(e){return e===t},u,!0),l=at(function(e){return T.call(t,e)>-1},u,!0),h=[function(e,n,r){return!o&&(r||n!==c)||((t=n).nodeType?f(e,n,r):l(e,n,r))}];for(;a<s;a++)if(n=i.relative[e[a].type])h=[at(ft(h),n)];else{n=i.filter[e[a].type].apply(null,e[a].matches);if(n[d]){r=++a;for(;r<s;r++)if(i.relative[e[r].type])break;return ct(a>1&&ft(h),a>1&&e.slice(0,a-1).join("").replace(j,"$1"),n,a<r&&ht(e.slice(a,r)),r<s&&ht(e=e.slice(r)),r<s&&e.join(""))}h.push(n)}return ft(h)}function pt(e,t){var r=t.length>0,s=e.length>0,o=function(u,a,f,l,h){var p,d,v,m=[],y=0,w="0",x=u&&[],T=h!=null,N=c,C=u||s&&i.find.TAG("*",h&&a.parentNode||a),k=b+=N==null?1:Math.E;T&&(c=a!==g&&a,n=o.el);for(;(p=C[w])!=null;w++){if(s&&p){for(d=0;v=e[d];d++)if(v(p,a,f)){l.push(p);break}T&&(b=k,n=++o.el)}r&&((p=!v&&p)&&y--,u&&x.push(p))}y+=w;if(r&&w!==y){for(d=0;v=t[d];d++)v(x,m,a,f);if(u){if(y>0)while(w--)!x[w]&&!m[w]&&(m[w]=E.call(l));m=lt(m)}S.apply(l,m),T&&!u&&m.length>0&&y+t.length>1&&nt.uniqueSort(l)}return T&&(b=k,c=N),x};return o.el=0,r?N(o):o}function dt(e,t,n){var r=0,i=t.length;for(;r<i;r++)nt(e,t[r],n);return n}function vt(e,t,n,r,s){var o,u,f,l,c,h=ut(e),p=h.length;if(!r&&h.length===1){u=h[0]=h[0].slice(0);if(u.length>2&&(f=u[0]).type==="ID"&&t.nodeType===9&&!s&&i.relative[u[1].type]){t=i.find.ID(f.matches[0].replace($,""),t,s)[0];if(!t)return n;e=e.slice(u.shift().length)}for(o=J.POS.test(e)?-1:u.length-1;o>=0;o--){f=u[o];if(i.relative[l=f.type])break;if(c=i.find[l])if(r=c(f.matches[0].replace($,""),z.test(u[0].type)&&t.parentNode||t,s)){u.splice(o,1),e=r.length&&u.join("");if(!e)return S.apply(n,x.call(r,0)),n;break}}}return a(e,h)(r,t,s,n,z.test(e)),n}function mt(){}var n,r,i,s,o,u,a,f,l,c,h=!0,p="undefined",d=("sizcache"+Math.random()).replace(".",""),m=String,g=e.document,y=g.documentElement,b=0,w=0,E=[].pop,S=[].push,x=[].slice,T=[].indexOf||function(e){var t=0,n=this.length;for(;t<n;t++)if(this[t]===e)return t;return-1},N=function(e,t){return e[d]=t==null||t,e},C=function(){var e={},t=[];return N(function(n,r){return t.push(n)>i.cacheLength&&delete e[t.shift()],e[n+" "]=r},e)},k=C(),L=C(),A=C(),O="[\\x20\\t\\r\\n\\f]",M="(?:\\\\.|[-\\w]|[^\\x00-\\xa0])+",_=M.replace("w","w#"),D="([*^$|!~]?=)",P="\\["+O+"*("+M+")"+O+"*(?:"+D+O+"*(?:(['\"])((?:\\\\.|[^\\\\])*?)\\3|("+_+")|)|)"+O+"*\\]",H=":("+M+")(?:\\((?:(['\"])((?:\\\\.|[^\\\\])*?)\\2|([^()[\\]]*|(?:(?:"+P+")|[^:]|\\\\.)*|.*))\\)|)",B=":(even|odd|eq|gt|lt|nth|first|last)(?:\\("+O+"*((?:-\\d)?\\d*)"+O+"*\\)|)(?=[^-]|$)",j=new RegExp("^"+O+"+|((?:^|[^\\\\])(?:\\\\.)*)"+O+"+$","g"),F=new RegExp("^"+O+"*,"+O+"*"),I=new RegExp("^"+O+"*([\\x20\\t\\r\\n\\f>+~])"+O+"*"),q=new RegExp(H),R=/^(?:#([\w\-]+)|(\w+)|\.([\w\-]+))$/,U=/^:not/,z=/[\x20\t\r\n\f]*[+~]/,W=/:not\($/,X=/h\d/i,V=/input|select|textarea|button/i,$=/\\(?!\\)/g,J={ID:new RegExp("^#("+M+")"),CLASS:new RegExp("^\\.("+M+")"),NAME:new RegExp("^\\[name=['\"]?("+M+")['\"]?\\]"),TAG:new RegExp("^("+M.replace("w","w*")+")"),ATTR:new RegExp("^"+P),PSEUDO:new RegExp("^"+H),POS:new RegExp(B,"i"),CHILD:new RegExp("^:(only|nth|first|last)-child(?:\\("+O+"*(even|odd|(([+-]|)(\\d*)n|)"+O+"*(?:([+-]|)"+O+"*(\\d+)|))"+O+"*\\)|)","i"),needsContext:new RegExp("^"+O+"*[>+~]|"+B,"i")},K=function(e){var t=g.createElement("div");try{return e(t)}catch(n){return!1}finally{t=null}},Q=K(function(e){return e.appendChild(g.createComment("")),!e.getElementsByTagName("*").length}),G=K(function(e){return e.innerHTML="<a href='#'></a>",e.firstChild&&typeof e.firstChild.getAttribute!==p&&e.firstChild.getAttribute("href")==="#"}),Y=K(function(e){e.innerHTML="<select></select>";var t=typeof e.lastChild.getAttribute("multiple");return t!=="boolean"&&t!=="string"}),Z=K(function(e){return e.innerHTML="<div class='hidden e'></div><div class='hidden'></div>",!e.getElementsByClassName||!e.getElementsByClassName("e").length?!1:(e.lastChild.className="e",e.getElementsByClassName("e").length===2)}),et=K(function(e){e.id=d+0,e.innerHTML="<a name='"+d+"'></a><div name='"+d+"'></div>",y.insertBefore(e,y.firstChild);var t=g.getElementsByName&&g.getElementsByName(d).length===2+g.getElementsByName(d+0).length;return r=!g.getElementById(d),y.removeChild(e),t});try{x.call(y.childNodes,0)[0].nodeType}catch(tt){x=function(e){var t,n=[];for(;t=this[e];e++)n.push(t);return n}}nt.matches=function(e,t){return nt(e,null,null,t)},nt.matchesSelector=function(e,t){return nt(t,null,null,[e]).length>0},s=nt.getText=function(e){var t,n="",r=0,i=e.nodeType;if(i){if(i===1||i===9||i===11){if(typeof e.textContent=="string")return e.textContent;for(e=e.firstChild;e;e=e.nextSibling)n+=s(e)}else if(i===3||i===4)return e.nodeValue}else for(;t=e[r];r++)n+=s(t);return n},o=nt.isXML=function(e){var t=e&&(e.ownerDocument||e).documentElement;return t?t.nodeName!=="HTML":!1},u=nt.contains=y.contains?function(e,t){var n=e.nodeType===9?e.documentElement:e,r=t&&t.parentNode;return e===r||!!(r&&r.nodeType===1&&n.contains&&n.contains(r))}:y.compareDocumentPosition?function(e,t){return t&&!!(e.compareDocumentPosition(t)&16)}:function(e,t){while(t=t.parentNode)if(t===e)return!0;return!1},nt.attr=function(e,t){var n,r=o(e);return r||(t=t.toLowerCase()),(n=i.attrHandle[t])?n(e):r||Y?e.getAttribute(t):(n=e.getAttributeNode(t),n?typeof e[t]=="boolean"?e[t]?t:null:n.specified?n.value:null:null)},i=nt.selectors={cacheLength:50,createPseudo:N,match:J,attrHandle:G?{}:{href:function(e){return e.getAttribute("href",2)},type:function(e){return e.getAttribute("type")}},find:{ID:r?function(e,t,n){if(typeof t.getElementById!==p&&!n){var r=t.getElementById(e);return r&&r.parentNode?[r]:[]}}:function(e,n,r){if(typeof n.getElementById!==p&&!r){var i=n.getElementById(e);return i?i.id===e||typeof i.getAttributeNode!==p&&i.getAttributeNode("id").value===e?[i]:t:[]}},TAG:Q?function(e,t){if(typeof t.getElementsByTagName!==p)return t.getElementsByTagName(e)}:function(e,t){var n=t.getElementsByTagName(e);if(e==="*"){var r,i=[],s=0;for(;r=n[s];s++)r.nodeType===1&&i.push(r);return i}return n},NAME:et&&function(e,t){if(typeof t.getElementsByName!==p)return t.getElementsByName(name)},CLASS:Z&&function(e,t,n){if(typeof t.getElementsByClassName!==p&&!n)return t.getElementsByClassName(e)}},relative:{">":{dir:"parentNode",first:!0}," ":{dir:"parentNode"},"+":{dir:"previousSibling",first:!0},"~":{dir:"previousSibling"}},preFilter:{ATTR:function(e){return e[1]=e[1].replace($,""),e[3]=(e[4]||e[5]||"").replace($,""),e[2]==="~="&&(e[3]=" "+e[3]+" "),e.slice(0,4)},CHILD:function(e){return e[1]=e[1].toLowerCase(),e[1]==="nth"?(e[2]||nt.error(e[0]),e[3]=+(e[3]?e[4]+(e[5]||1):2*(e[2]==="even"||e[2]==="odd")),e[4]=+(e[6]+e[7]||e[2]==="odd")):e[2]&&nt.error(e[0]),e},PSEUDO:function(e){var t,n;if(J.CHILD.test(e[0]))return null;if(e[3])e[2]=e[3];else if(t=e[4])q.test(t)&&(n=ut(t,!0))&&(n=t.indexOf(")",t.length-n)-t.length)&&(t=t.slice(0,n),e[0]=e[0].slice(0,n)),e[2]=t;return e.slice(0,3)}},filter:{ID:r?function(e){return e=e.replace($,""),function(t){return t.getAttribute("id")===e}}:function(e){return e=e.replace($,""),function(t){var n=typeof t.getAttributeNode!==p&&t.getAttributeNode("id");return n&&n.value===e}},TAG:function(e){return e==="*"?function(){return!0}:(e=e.replace($,"").toLowerCase(),function(t){return t.nodeName&&t.nodeName.toLowerCase()===e})},CLASS:function(e){var t=k[d][e+" "];return t||(t=new RegExp("(^|"+O+")"+e+"("+O+"|$)"))&&k(e,function(e){return t.test(e.className||typeof e.getAttribute!==p&&e.getAttribute("class")||"")})},ATTR:function(e,t,n){return function(r,i){var s=nt.attr(r,e);return s==null?t==="!=":t?(s+="",t==="="?s===n:t==="!="?s!==n:t==="^="?n&&s.indexOf(n)===0:t==="*="?n&&s.indexOf(n)>-1:t==="$="?n&&s.substr(s.length-n.length)===n:t==="~="?(" "+s+" ").indexOf(n)>-1:t==="|="?s===n||s.substr(0,n.length+1)===n+"-":!1):!0}},CHILD:function(e,t,n,r){return e==="nth"?function(e){var t,i,s=e.parentNode;if(n===1&&r===0)return!0;if(s){i=0;for(t=s.firstChild;t;t=t.nextSibling)if(t.nodeType===1){i++;if(e===t)break}}return i-=r,i===n||i%n===0&&i/n>=0}:function(t){var n=t;switch(e){case"only":case"first":while(n=n.previousSibling)if(n.nodeType===1)return!1;if(e==="first")return!0;n=t;case"last":while(n=n.nextSibling)if(n.nodeType===1)return!1;return!0}}},PSEUDO:function(e,t){var n,r=i.pseudos[e]||i.setFilters[e.toLowerCase()]||nt.error("unsupported pseudo: "+e);return r[d]?r(t):r.length>1?(n=[e,e,"",t],i.setFilters.hasOwnProperty(e.toLowerCase())?N(function(e,n){var i,s=r(e,t),o=s.length;while(o--)i=T.call(e,s[o]),e[i]=!(n[i]=s[o])}):function(e){return r(e,0,n)}):r}},pseudos:{not:N(function(e){var t=[],n=[],r=a(e.replace(j,"$1"));return r[d]?N(function(e,t,n,i){var s,o=r(e,null,i,[]),u=e.length;while(u--)if(s=o[u])e[u]=!(t[u]=s)}):function(e,i,s){return t[0]=e,r(t,null,s,n),!n.pop()}}),has:N(function(e){return function(t){return nt(e,t).length>0}}),contains:N(function(e){return function(t){return(t.textContent||t.innerText||s(t)).indexOf(e)>-1}}),enabled:function(e){return e.disabled===!1},disabled:function(e){return e.disabled===!0},checked:function(e){var t=e.nodeName.toLowerCase();return t==="input"&&!!e.checked||t==="option"&&!!e.selected},selected:function(e){return e.parentNode&&e.parentNode.selectedIndex,e.selected===!0},parent:function(e){return!i.pseudos.empty(e)},empty:function(e){var t;e=e.firstChild;while(e){if(e.nodeName>"@"||(t=e.nodeType)===3||t===4)return!1;e=e.nextSibling}return!0},header:function(e){return X.test(e.nodeName)},text:function(e){var t,n;return e.nodeName.toLowerCase()==="input"&&(t=e.type)==="text"&&((n=e.getAttribute("type"))==null||n.toLowerCase()===t)},radio:rt("radio"),checkbox:rt("checkbox"),file:rt("file"),password:rt("password"),image:rt("image"),submit:it("submit"),reset:it("reset"),button:function(e){var t=e.nodeName.toLowerCase();return t==="input"&&e.type==="button"||t==="button"},input:function(e){return V.test(e.nodeName)},focus:function(e){var t=e.ownerDocument;return e===t.activeElement&&(!t.hasFocus||t.hasFocus())&&!!(e.type||e.href||~e.tabIndex)},active:function(e){return e===e.ownerDocument.activeElement},first:st(function(){return[0]}),last:st(function(e,t){return[t-1]}),eq:st(function(e,t,n){return[n<0?n+t:n]}),even:st(function(e,t){for(var n=0;n<t;n+=2)e.push(n);return e}),odd:st(function(e,t){for(var n=1;n<t;n+=2)e.push(n);return e}),lt:st(function(e,t,n){for(var r=n<0?n+t:n;--r>=0;)e.push(r);return e}),gt:st(function(e,t,n){for(var r=n<0?n+t:n;++r<t;)e.push(r);return e})}},f=y.compareDocumentPosition?function(e,t){return e===t?(l=!0,0):(!e.compareDocumentPosition||!t.compareDocumentPosition?e.compareDocumentPosition:e.compareDocumentPosition(t)&4)?-1:1}:function(e,t){if(e===t)return l=!0,0;if(e.sourceIndex&&t.sourceIndex)return e.sourceIndex-t.sourceIndex;var n,r,i=[],s=[],o=e.parentNode,u=t.parentNode,a=o;if(o===u)return ot(e,t);if(!o)return-1;if(!u)return 1;while(a)i.unshift(a),a=a.parentNode;a=u;while(a)s.unshift(a),a=a.parentNode;n=i.length,r=s.length;for(var f=0;f<n&&f<r;f++)if(i[f]!==s[f])return ot(i[f],s[f]);return f===n?ot(e,s[f],-1):ot(i[f],t,1)},[0,0].sort(f),h=!l,nt.uniqueSort=function(e){var t,n=[],r=1,i=0;l=h,e.sort(f);if(l){for(;t=e[r];r++)t===e[r-1]&&(i=n.push(r));while(i--)e.splice(n[i],1)}return e},nt.error=function(e){throw new Error("Syntax error, unrecognized expression: "+e)},a=nt.compile=function(e,t){var n,r=[],i=[],s=A[d][e+" "];if(!s){t||(t=ut(e)),n=t.length;while(n--)s=ht(t[n]),s[d]?r.push(s):i.push(s);s=A(e,pt(i,r))}return s},g.querySelectorAll&&function(){var e,t=vt,n=/'|\\/g,r=/\=[\x20\t\r\n\f]*([^'"\]]*)[\x20\t\r\n\f]*\]/g,i=[":focus"],s=[":active"],u=y.matchesSelector||y.mozMatchesSelector||y.webkitMatchesSelector||y.oMatchesSelector||y.msMatchesSelector;K(function(e){e.innerHTML="<select><option selected=''></option></select>",e.querySelectorAll("[selected]").length||i.push("\\["+O+"*(?:checked|disabled|ismap|multiple|readonly|selected|value)"),e.querySelectorAll(":checked").length||i.push(":checked")}),K(function(e){e.innerHTML="<p test=''></p>",e.querySelectorAll("[test^='']").length&&i.push("[*^$]="+O+"*(?:\"\"|'')"),e.innerHTML="<input type='hidden'/>",e.querySelectorAll(":enabled").length||i.push(":enabled",":disabled")}),i=new RegExp(i.join("|")),vt=function(e,r,s,o,u){if(!o&&!u&&!i.test(e)){var a,f,l=!0,c=d,h=r,p=r.nodeType===9&&e;if(r.nodeType===1&&r.nodeName.toLowerCase()!=="object"){a=ut(e),(l=r.getAttribute("id"))?c=l.replace(n,"\\$&"):r.setAttribute("id",c),c="[id='"+c+"'] ",f=a.length;while(f--)a[f]=c+a[f].join("");h=z.test(e)&&r.parentNode||r,p=a.join(",")}if(p)try{return S.apply(s,x.call(h.querySelectorAll(p),0)),s}catch(v){}finally{l||r.removeAttribute("id")}}return t(e,r,s,o,u)},u&&(K(function(t){e=u.call(t,"div");try{u.call(t,"[test!='']:sizzle"),s.push("!=",H)}catch(n){}}),s=new RegExp(s.join("|")),nt.matchesSelector=function(t,n){n=n.replace(r,"='$1']");if(!o(t)&&!s.test(n)&&!i.test(n))try{var a=u.call(t,n);if(a||e||t.document&&t.document.nodeType!==11)return a}catch(f){}return nt(n,null,null,[t]).length>0})}(),i.pseudos.nth=i.pseudos.eq,i.filters=mt.prototype=i.pseudos,i.setFilters=new mt,nt.attr=v.attr,v.find=nt,v.expr=nt.selectors,v.expr[":"]=v.expr.pseudos,v.unique=nt.uniqueSort,v.text=nt.getText,v.isXMLDoc=nt.isXML,v.contains=nt.contains}(e);var nt=/Until$/,rt=/^(?:parents|prev(?:Until|All))/,it=/^.[^:#\[\.,]*$/,st=v.expr.match.needsContext,ot={children:!0,contents:!0,next:!0,prev:!0};v.fn.extend({find:function(e){var t,n,r,i,s,o,u=this;if(typeof e!="string")return v(e).filter(function(){for(t=0,n=u.length;t<n;t++)if(v.contains(u[t],this))return!0});o=this.pushStack("","find",e);for(t=0,n=this.length;t<n;t++){r=o.length,v.find(e,this[t],o);if(t>0)for(i=r;i<o.length;i++)for(s=0;s<r;s++)if(o[s]===o[i]){o.splice(i--,1);break}}return o},has:function(e){var t,n=v(e,this),r=n.length;return this.filter(function(){for(t=0;t<r;t++)if(v.contains(this,n[t]))return!0})},not:function(e){return this.pushStack(ft(this,e,!1),"not",e)},filter:function(e){return this.pushStack(ft(this,e,!0),"filter",e)},is:function(e){return!!e&&(typeof e=="string"?st.test(e)?v(e,this.context).index(this[0])>=0:v.filter(e,this).length>0:this.filter(e).length>0)},closest:function(e,t){var n,r=0,i=this.length,s=[],o=st.test(e)||typeof e!="string"?v(e,t||this.context):0;for(;r<i;r++){n=this[r];while(n&&n.ownerDocument&&n!==t&&n.nodeType!==11){if(o?o.index(n)>-1:v.find.matchesSelector(n,e)){s.push(n);break}n=n.parentNode}}return s=s.length>1?v.unique(s):s,this.pushStack(s,"closest",e)},index:function(e){return e?typeof e=="string"?v.inArray(this[0],v(e)):v.inArray(e.jquery?e[0]:e,this):this[0]&&this[0].parentNode?this.prevAll().length:-1},add:function(e,t){var n=typeof e=="string"?v(e,t):v.makeArray(e&&e.nodeType?[e]:e),r=v.merge(this.get(),n);return this.pushStack(ut(n[0])||ut(r[0])?r:v.unique(r))},addBack:function(e){return this.add(e==null?this.prevObject:this.prevObject.filter(e))}}),v.fn.andSelf=v.fn.addBack,v.each({parent:function(e){var t=e.parentNode;return t&&t.nodeType!==11?t:null},parents:function(e){return v.dir(e,"parentNode")},parentsUntil:function(e,t,n){return v.dir(e,"parentNode",n)},next:function(e){return at(e,"nextSibling")},prev:function(e){return at(e,"previousSibling")},nextAll:function(e){return v.dir(e,"nextSibling")},prevAll:function(e){return v.dir(e,"previousSibling")},nextUntil:function(e,t,n){return v.dir(e,"nextSibling",n)},prevUntil:function(e,t,n){return v.dir(e,"previousSibling",n)},siblings:function(e){return v.sibling((e.parentNode||{}).firstChild,e)},children:function(e){return v.sibling(e.firstChild)},contents:function(e){return v.nodeName(e,"iframe")?e.contentDocument||e.contentWindow.document:v.merge([],e.childNodes)}},function(e,t){v.fn[e]=function(n,r){var i=v.map(this,t,n);return nt.test(e)||(r=n),r&&typeof r=="string"&&(i=v.filter(r,i)),i=this.length>1&&!ot[e]?v.unique(i):i,this.length>1&&rt.test(e)&&(i=i.reverse()),this.pushStack(i,e,l.call(arguments).join(","))}}),v.extend({filter:function(e,t,n){return n&&(e=":not("+e+")"),t.length===1?v.find.matchesSelector(t[0],e)?[t[0]]:[]:v.find.matches(e,t)},dir:function(e,n,r){var i=[],s=e[n];while(s&&s.nodeType!==9&&(r===t||s.nodeType!==1||!v(s).is(r)))s.nodeType===1&&i.push(s),s=s[n];return i},sibling:function(e,t){var n=[];for(;e;e=e.nextSibling)e.nodeType===1&&e!==t&&n.push(e);return n}});var ct="abbr|article|aside|audio|bdi|canvas|data|datalist|details|figcaption|figure|footer|header|hgroup|mark|meter|nav|output|progress|section|summary|time|video",ht=/ jQuery\d+="(?:null|\d+)"/g,pt=/^\s+/,dt=/<(?!area|br|col|embed|hr|img|input|link|meta|param)(([\w:]+)[^>]*)\/>/gi,vt=/<([\w:]+)/,mt=/<tbody/i,gt=/<|&#?\w+;/,yt=/<(?:script|style|link)/i,bt=/<(?:script|object|embed|option|style)/i,wt=new RegExp("<(?:"+ct+")[\\s/>]","i"),Et=/^(?:checkbox|radio)$/,St=/checked\s*(?:[^=]|=\s*.checked.)/i,xt=/\/(java|ecma)script/i,Tt=/^\s*<!(?:\[CDATA\[|\-\-)|[\]\-]{2}>\s*$/g,Nt={option:[1,"<select multiple='multiple'>","</select>"],legend:[1,"<fieldset>","</fieldset>"],thead:[1,"<table>","</table>"],tr:[2,"<table><tbody>","</tbody></table>"],td:[3,"<table><tbody><tr>","</tr></tbody></table>"],col:[2,"<table><tbody></tbody><colgroup>","</colgroup></table>"],area:[1,"<map>","</map>"],_default:[0,"",""]},Ct=lt(i),kt=Ct.appendChild(i.createElement("div"));Nt.optgroup=Nt.option,Nt.tbody=Nt.tfoot=Nt.colgroup=Nt.caption=Nt.thead,Nt.th=Nt.td,v.support.htmlSerialize||(Nt._default=[1,"X<div>","</div>"]),v.fn.extend({text:function(e){return v.access(this,function(e){return e===t?v.text(this):this.empty().append((this[0]&&this[0].ownerDocument||i).createTextNode(e))},null,e,arguments.length)},wrapAll:function(e){if(v.isFunction(e))return this.each(function(t){v(this).wrapAll(e.call(this,t))});if(this[0]){var t=v(e,this[0].ownerDocument).eq(0).clone(!0);this[0].parentNode&&t.insertBefore(this[0]),t.map(function(){var e=this;while(e.firstChild&&e.firstChild.nodeType===1)e=e.firstChild;return e}).append(this)}return this},wrapInner:function(e){return v.isFunction(e)?this.each(function(t){v(this).wrapInner(e.call(this,t))}):this.each(function(){var t=v(this),n=t.contents();n.length?n.wrapAll(e):t.append(e)})},wrap:function(e){var t=v.isFunction(e);return this.each(function(n){v(this).wrapAll(t?e.call(this,n):e)})},unwrap:function(){return this.parent().each(function(){v.nodeName(this,"body")||v(this).replaceWith(this.childNodes)}).end()},append:function(){return this.domManip(arguments,!0,function(e){(this.nodeType===1||this.nodeType===11)&&this.appendChild(e)})},prepend:function(){return this.domManip(arguments,!0,function(e){(this.nodeType===1||this.nodeType===11)&&this.insertBefore(e,this.firstChild)})},before:function(){if(!ut(this[0]))return this.domManip(arguments,!1,function(e){this.parentNode.insertBefore(e,this)});if(arguments.length){var e=v.clean(arguments);return this.pushStack(v.merge(e,this),"before",this.selector)}},after:function(){if(!ut(this[0]))return this.domManip(arguments,!1,function(e){this.parentNode.insertBefore(e,this.nextSibling)});if(arguments.length){var e=v.clean(arguments);return this.pushStack(v.merge(this,e),"after",this.selector)}},remove:function(e,t){var n,r=0;for(;(n=this[r])!=null;r++)if(!e||v.filter(e,[n]).length)!t&&n.nodeType===1&&(v.cleanData(n.getElementsByTagName("*")),v.cleanData([n])),n.parentNode&&n.parentNode.removeChild(n);return this},empty:function(){var e,t=0;for(;(e=this[t])!=null;t++){e.nodeType===1&&v.cleanData(e.getElementsByTagName("*"));while(e.firstChild)e.removeChild(e.firstChild)}return this},clone:function(e,t){return e=e==null?!1:e,t=t==null?e:t,this.map(function(){return v.clone(this,e,t)})},html:function(e){return v.access(this,function(e){var n=this[0]||{},r=0,i=this.length;if(e===t)return n.nodeType===1?n.innerHTML.replace(ht,""):t;if(typeof e=="string"&&!yt.test(e)&&(v.support.htmlSerialize||!wt.test(e))&&(v.support.leadingWhitespace||!pt.test(e))&&!Nt[(vt.exec(e)||["",""])[1].toLowerCase()]){e=e.replace(dt,"<$1></$2>");try{for(;r<i;r++)n=this[r]||{},n.nodeType===1&&(v.cleanData(n.getElementsByTagName("*")),n.innerHTML=e);n=0}catch(s){}}n&&this.empty().append(e)},null,e,arguments.length)},replaceWith:function(e){return ut(this[0])?this.length?this.pushStack(v(v.isFunction(e)?e():e),"replaceWith",e):this:v.isFunction(e)?this.each(function(t){var n=v(this),r=n.html();n.replaceWith(e.call(this,t,r))}):(typeof e!="string"&&(e=v(e).detach()),this.each(function(){var t=this.nextSibling,n=this.parentNode;v(this).remove(),t?v(t).before(e):v(n).append(e)}))},detach:function(e){return this.remove(e,!0)},domManip:function(e,n,r){e=[].concat.apply([],e);var i,s,o,u,a=0,f=e[0],l=[],c=this.length;if(!v.support.checkClone&&c>1&&typeof f=="string"&&St.test(f))return this.each(function(){v(this).domManip(e,n,r)});if(v.isFunction(f))return this.each(function(i){var s=v(this);e[0]=f.call(this,i,n?s.html():t),s.domManip(e,n,r)});if(this[0]){i=v.buildFragment(e,this,l),o=i.fragment,s=o.firstChild,o.childNodes.length===1&&(o=s);if(s){n=n&&v.nodeName(s,"tr");for(u=i.cacheable||c-1;a<c;a++)r.call(n&&v.nodeName(this[a],"table")?Lt(this[a],"tbody"):this[a],a===u?o:v.clone(o,!0,!0))}o=s=null,l.length&&v.each(l,function(e,t){t.src?v.ajax?v.ajax({url:t.src,type:"GET",dataType:"script",async:!1,global:!1,"throws":!0}):v.error("no ajax"):v.globalEval((t.text||t.textContent||t.innerHTML||"").replace(Tt,"")),t.parentNode&&t.parentNode.removeChild(t)})}return this}}),v.buildFragment=function(e,n,r){var s,o,u,a=e[0];return n=n||i,n=!n.nodeType&&n[0]||n,n=n.ownerDocument||n,e.length===1&&typeof a=="string"&&a.length<512&&n===i&&a.charAt(0)==="<"&&!bt.test(a)&&(v.support.checkClone||!St.test(a))&&(v.support.html5Clone||!wt.test(a))&&(o=!0,s=v.fragments[a],u=s!==t),s||(s=n.createDocumentFragment(),v.clean(e,n,s,r),o&&(v.fragments[a]=u&&s)),{fragment:s,cacheable:o}},v.fragments={},v.each({appendTo:"append",prependTo:"prepend",insertBefore:"before",insertAfter:"after",replaceAll:"replaceWith"},function(e,t){v.fn[e]=function(n){var r,i=0,s=[],o=v(n),u=o.length,a=this.length===1&&this[0].parentNode;if((a==null||a&&a.nodeType===11&&a.childNodes.length===1)&&u===1)return o[t](this[0]),this;for(;i<u;i++)r=(i>0?this.clone(!0):this).get(),v(o[i])[t](r),s=s.concat(r);return this.pushStack(s,e,o.selector)}}),v.extend({clone:function(e,t,n){var r,i,s,o;v.support.html5Clone||v.isXMLDoc(e)||!wt.test("<"+e.nodeName+">")?o=e.cloneNode(!0):(kt.innerHTML=e.outerHTML,kt.removeChild(o=kt.firstChild));if((!v.support.noCloneEvent||!v.support.noCloneChecked)&&(e.nodeType===1||e.nodeType===11)&&!v.isXMLDoc(e)){Ot(e,o),r=Mt(e),i=Mt(o);for(s=0;r[s];++s)i[s]&&Ot(r[s],i[s])}if(t){At(e,o);if(n){r=Mt(e),i=Mt(o);for(s=0;r[s];++s)At(r[s],i[s])}}return r=i=null,o},clean:function(e,t,n,r){var s,o,u,a,f,l,c,h,p,d,m,g,y=t===i&&Ct,b=[];if(!t||typeof t.createDocumentFragment=="undefined")t=i;for(s=0;(u=e[s])!=null;s++){typeof u=="number"&&(u+="");if(!u)continue;if(typeof u=="string")if(!gt.test(u))u=t.createTextNode(u);else{y=y||lt(t),c=t.createElement("div"),y.appendChild(c),u=u.replace(dt,"<$1></$2>"),a=(vt.exec(u)||["",""])[1].toLowerCase(),f=Nt[a]||Nt._default,l=f[0],c.innerHTML=f[1]+u+f[2];while(l--)c=c.lastChild;if(!v.support.tbody){h=mt.test(u),p=a==="table"&&!h?c.firstChild&&c.firstChild.childNodes:f[1]==="<table>"&&!h?c.childNodes:[];for(o=p.length-1;o>=0;--o)v.nodeName(p[o],"tbody")&&!p[o].childNodes.length&&p[o].parentNode.removeChild(p[o])}!v.support.leadingWhitespace&&pt.test(u)&&c.insertBefore(t.createTextNode(pt.exec(u)[0]),c.firstChild),u=c.childNodes,c.parentNode.removeChild(c)}u.nodeType?b.push(u):v.merge(b,u)}c&&(u=c=y=null);if(!v.support.appendChecked)for(s=0;(u=b[s])!=null;s++)v.nodeName(u,"input")?_t(u):typeof u.getElementsByTagName!="undefined"&&v.grep(u.getElementsByTagName("input"),_t);if(n){m=function(e){if(!e.type||xt.test(e.type))return r?r.push(e.parentNode?e.parentNode.removeChild(e):e):n.appendChild(e)};for(s=0;(u=b[s])!=null;s++)if(!v.nodeName(u,"script")||!m(u))n.appendChild(u),typeof u.getElementsByTagName!="undefined"&&(g=v.grep(v.merge([],u.getElementsByTagName("script")),m),b.splice.apply(b,[s+1,0].concat(g)),s+=g.length)}return b},cleanData:function(e,t){var n,r,i,s,o=0,u=v.expando,a=v.cache,f=v.support.deleteExpando,l=v.event.special;for(;(i=e[o])!=null;o++)if(t||v.acceptData(i)){r=i[u],n=r&&a[r];if(n){if(n.events)for(s in n.events)l[s]?v.event.remove(i,s):v.removeEvent(i,s,n.handle);a[r]&&(delete a[r],f?delete i[u]:i.removeAttribute?i.removeAttribute(u):i[u]=null,v.deletedIds.push(r))}}}}),function(){var e,t;v.uaMatch=function(e){e=e.toLowerCase();var t=/(chrome)[ \/]([\w.]+)/.exec(e)||/(webkit)[ \/]([\w.]+)/.exec(e)||/(opera)(?:.*version|)[ \/]([\w.]+)/.exec(e)||/(msie) ([\w.]+)/.exec(e)||e.indexOf("compatible")<0&&/(mozilla)(?:.*? rv:([\w.]+)|)/.exec(e)||[];return{browser:t[1]||"",version:t[2]||"0"}},e=v.uaMatch(o.userAgent),t={},e.browser&&(t[e.browser]=!0,t.version=e.version),t.chrome?t.webkit=!0:t.webkit&&(t.safari=!0),v.browser=t,v.sub=function(){function e(t,n){return new e.fn.init(t,n)}v.extend(!0,e,this),e.superclass=this,e.fn=e.prototype=this(),e.fn.constructor=e,e.sub=this.sub,e.fn.init=function(r,i){return i&&i instanceof v&&!(i instanceof e)&&(i=e(i)),v.fn.init.call(this,r,i,t)},e.fn.init.prototype=e.fn;var t=e(i);return e}}();var Dt,Pt,Ht,Bt=/alpha\([^)]*\)/i,jt=/opacity=([^)]*)/,Ft=/^(top|right|bottom|left)$/,It=/^(none|table(?!-c[ea]).+)/,qt=/^margin/,Rt=new RegExp("^("+m+")(.*)$","i"),Ut=new RegExp("^("+m+")(?!px)[a-z%]+$","i"),zt=new RegExp("^([-+])=("+m+")","i"),Wt={BODY:"block"},Xt={position:"absolute",visibility:"hidden",display:"block"},Vt={letterSpacing:0,fontWeight:400},$t=["Top","Right","Bottom","Left"],Jt=["Webkit","O","Moz","ms"],Kt=v.fn.toggle;v.fn.extend({css:function(e,n){return v.access(this,function(e,n,r){return r!==t?v.style(e,n,r):v.css(e,n)},e,n,arguments.length>1)},show:function(){return Yt(this,!0)},hide:function(){return Yt(this)},toggle:function(e,t){var n=typeof e=="boolean";return v.isFunction(e)&&v.isFunction(t)?Kt.apply(this,arguments):this.each(function(){(n?e:Gt(this))?v(this).show():v(this).hide()})}}),v.extend({cssHooks:{opacity:{get:function(e,t){if(t){var n=Dt(e,"opacity");return n===""?"1":n}}}},cssNumber:{fillOpacity:!0,fontWeight:!0,lineHeight:!0,opacity:!0,orphans:!0,widows:!0,zIndex:!0,zoom:!0},cssProps:{"float":v.support.cssFloat?"cssFloat":"styleFloat"},style:function(e,n,r,i){if(!e||e.nodeType===3||e.nodeType===8||!e.style)return;var s,o,u,a=v.camelCase(n),f=e.style;n=v.cssProps[a]||(v.cssProps[a]=Qt(f,a)),u=v.cssHooks[n]||v.cssHooks[a];if(r===t)return u&&"get"in u&&(s=u.get(e,!1,i))!==t?s:f[n];o=typeof r,o==="string"&&(s=zt.exec(r))&&(r=(s[1]+1)*s[2]+parseFloat(v.css(e,n)),o="number");if(r==null||o==="number"&&isNaN(r))return;o==="number"&&!v.cssNumber[a]&&(r+="px");if(!u||!("set"in u)||(r=u.set(e,r,i))!==t)try{f[n]=r}catch(l){}},css:function(e,n,r,i){var s,o,u,a=v.camelCase(n);return n=v.cssProps[a]||(v.cssProps[a]=Qt(e.style,a)),u=v.cssHooks[n]||v.cssHooks[a],u&&"get"in u&&(s=u.get(e,!0,i)),s===t&&(s=Dt(e,n)),s==="normal"&&n in Vt&&(s=Vt[n]),r||i!==t?(o=parseFloat(s),r||v.isNumeric(o)?o||0:s):s},swap:function(e,t,n){var r,i,s={};for(i in t)s[i]=e.style[i],e.style[i]=t[i];r=n.call(e);for(i in t)e.style[i]=s[i];return r}}),e.getComputedStyle?Dt=function(t,n){var r,i,s,o,u=e.getComputedStyle(t,null),a=t.style;return u&&(r=u.getPropertyValue(n)||u[n],r===""&&!v.contains(t.ownerDocument,t)&&(r=v.style(t,n)),Ut.test(r)&&qt.test(n)&&(i=a.width,s=a.minWidth,o=a.maxWidth,a.minWidth=a.maxWidth=a.width=r,r=u.width,a.width=i,a.minWidth=s,a.maxWidth=o)),r}:i.documentElement.currentStyle&&(Dt=function(e,t){var n,r,i=e.currentStyle&&e.currentStyle[t],s=e.style;return i==null&&s&&s[t]&&(i=s[t]),Ut.test(i)&&!Ft.test(t)&&(n=s.left,r=e.runtimeStyle&&e.runtimeStyle.left,r&&(e.runtimeStyle.left=e.currentStyle.left),s.left=t==="fontSize"?"1em":i,i=s.pixelLeft+"px",s.left=n,r&&(e.runtimeStyle.left=r)),i===""?"auto":i}),v.each(["height","width"],function(e,t){v.cssHooks[t]={get:function(e,n,r){if(n)return e.offsetWidth===0&&It.test(Dt(e,"display"))?v.swap(e,Xt,function(){return tn(e,t,r)}):tn(e,t,r)},set:function(e,n,r){return Zt(e,n,r?en(e,t,r,v.support.boxSizing&&v.css(e,"boxSizing")==="border-box"):0)}}}),v.support.opacity||(v.cssHooks.opacity={get:function(e,t){return jt.test((t&&e.currentStyle?e.currentStyle.filter:e.style.filter)||"")?.01*parseFloat(RegExp.$1)+"":t?"1":""},set:function(e,t){var n=e.style,r=e.currentStyle,i=v.isNumeric(t)?"alpha(opacity="+t*100+")":"",s=r&&r.filter||n.filter||"";n.zoom=1;if(t>=1&&v.trim(s.replace(Bt,""))===""&&n.removeAttribute){n.removeAttribute("filter");if(r&&!r.filter)return}n.filter=Bt.test(s)?s.replace(Bt,i):s+" "+i}}),v(function(){v.support.reliableMarginRight||(v.cssHooks.marginRight={get:function(e,t){return v.swap(e,{display:"inline-block"},function(){if(t)return Dt(e,"marginRight")})}}),!v.support.pixelPosition&&v.fn.position&&v.each(["top","left"],function(e,t){v.cssHooks[t]={get:function(e,n){if(n){var r=Dt(e,t);return Ut.test(r)?v(e).position()[t]+"px":r}}}})}),v.expr&&v.expr.filters&&(v.expr.filters.hidden=function(e){return e.offsetWidth===0&&e.offsetHeight===0||!v.support.reliableHiddenOffsets&&(e.style&&e.style.display||Dt(e,"display"))==="none"},v.expr.filters.visible=function(e){return!v.expr.filters.hidden(e)}),v.each({margin:"",padding:"",border:"Width"},function(e,t){v.cssHooks[e+t]={expand:function(n){var r,i=typeof n=="string"?n.split(" "):[n],s={};for(r=0;r<4;r++)s[e+$t[r]+t]=i[r]||i[r-2]||i[0];return s}},qt.test(e)||(v.cssHooks[e+t].set=Zt)});var rn=/%20/g,sn=/\[\]$/,on=/\r?\n/g,un=/^(?:color|date|datetime|datetime-local|email|hidden|month|number|password|range|search|tel|text|time|url|week)$/i,an=/^(?:select|textarea)/i;v.fn.extend({serialize:function(){return v.param(this.serializeArray())},serializeArray:function(){return this.map(function(){return this.elements?v.makeArray(this.elements):this}).filter(function(){return this.name&&!this.disabled&&(this.checked||an.test(this.nodeName)||un.test(this.type))}).map(function(e,t){var n=v(this).val();return n==null?null:v.isArray(n)?v.map(n,function(e,n){return{name:t.name,value:e.replace(on,"\r\n")}}):{name:t.name,value:n.replace(on,"\r\n")}}).get()}}),v.param=function(e,n){var r,i=[],s=function(e,t){t=v.isFunction(t)?t():t==null?"":t,i[i.length]=encodeURIComponent(e)+"="+encodeURIComponent(t)};n===t&&(n=v.ajaxSettings&&v.ajaxSettings.traditional);if(v.isArray(e)||e.jquery&&!v.isPlainObject(e))v.each(e,function(){s(this.name,this.value)});else for(r in e)fn(r,e[r],n,s);return i.join("&").replace(rn,"+")};var ln,cn,hn=/#.*$/,pn=/^(.*?):[ \t]*([^\r\n]*)\r?$/mg,dn=/^(?:about|app|app\-storage|.+\-extension|file|res|widget):$/,vn=/^(?:GET|HEAD)$/,mn=/^\/\//,gn=/\?/,yn=/<script\b[^<]*(?:(?!<\/script>)<[^<]*)*<\/script>/gi,bn=/([?&])_=[^&]*/,wn=/^([\w\+\.\-]+:)(?:\/\/([^\/?#:]*)(?::(\d+)|)|)/,En=v.fn.load,Sn={},xn={},Tn=["*/"]+["*"];try{cn=s.href}catch(Nn){cn=i.createElement("a"),cn.href="",cn=cn.href}ln=wn.exec(cn.toLowerCase())||[],v.fn.load=function(e,n,r){if(typeof e!="string"&&En)return En.apply(this,arguments);if(!this.length)return this;var i,s,o,u=this,a=e.indexOf(" ");return a>=0&&(i=e.slice(a,e.length),e=e.slice(0,a)),v.isFunction(n)?(r=n,n=t):n&&typeof n=="object"&&(s="POST"),v.ajax({url:e,type:s,dataType:"html",data:n,complete:function(e,t){r&&u.each(r,o||[e.responseText,t,e])}}).done(function(e){o=arguments,u.html(i?v("<div>").append(e.replace(yn,"")).find(i):e)}),this},v.each("ajaxStart ajaxStop ajaxComplete ajaxError ajaxSuccess ajaxSend".split(" "),function(e,t){v.fn[t]=function(e){return this.on(t,e)}}),v.each(["get","post"],function(e,n){v[n]=function(e,r,i,s){return v.isFunction(r)&&(s=s||i,i=r,r=t),v.ajax({type:n,url:e,data:r,success:i,dataType:s})}}),v.extend({getScript:function(e,n){return v.get(e,t,n,"script")},getJSON:function(e,t,n){return v.get(e,t,n,"json")},ajaxSetup:function(e,t){return t?Ln(e,v.ajaxSettings):(t=e,e=v.ajaxSettings),Ln(e,t),e},ajaxSettings:{url:cn,isLocal:dn.test(ln[1]),global:!0,type:"GET",contentType:"application/x-www-form-urlencoded; charset=UTF-8",processData:!0,async:!0,accepts:{xml:"application/xml, text/xml",html:"text/html",text:"text/plain",json:"application/json, text/javascript","*":Tn},contents:{xml:/xml/,html:/html/,json:/json/},responseFields:{xml:"responseXML",text:"responseText"},converters:{"* text":e.String,"text html":!0,"text json":v.parseJSON,"text xml":v.parseXML},flatOptions:{context:!0,url:!0}},ajaxPrefilter:Cn(Sn),ajaxTransport:Cn(xn),ajax:function(e,n){function T(e,n,s,a){var l,y,b,w,S,T=n;if(E===2)return;E=2,u&&clearTimeout(u),o=t,i=a||"",x.readyState=e>0?4:0,s&&(w=An(c,x,s));if(e>=200&&e<300||e===304)c.ifModified&&(S=x.getResponseHeader("Last-Modified"),S&&(v.lastModified[r]=S),S=x.getResponseHeader("Etag"),S&&(v.etag[r]=S)),e===304?(T="notmodified",l=!0):(l=On(c,w),T=l.state,y=l.data,b=l.error,l=!b);else{b=T;if(!T||e)T="error",e<0&&(e=0)}x.status=e,x.statusText=(n||T)+"",l?d.resolveWith(h,[y,T,x]):d.rejectWith(h,[x,T,b]),x.statusCode(g),g=t,f&&p.trigger("ajax"+(l?"Success":"Error"),[x,c,l?y:b]),m.fireWith(h,[x,T]),f&&(p.trigger("ajaxComplete",[x,c]),--v.active||v.event.trigger("ajaxStop"))}typeof e=="object"&&(n=e,e=t),n=n||{};var r,i,s,o,u,a,f,l,c=v.ajaxSetup({},n),h=c.context||c,p=h!==c&&(h.nodeType||h instanceof v)?v(h):v.event,d=v.Deferred(),m=v.Callbacks("once memory"),g=c.statusCode||{},b={},w={},E=0,S="canceled",x={readyState:0,setRequestHeader:function(e,t){if(!E){var n=e.toLowerCase();e=w[n]=w[n]||e,b[e]=t}return this},getAllResponseHeaders:function(){return E===2?i:null},getResponseHeader:function(e){var n;if(E===2){if(!s){s={};while(n=pn.exec(i))s[n[1].toLowerCase()]=n[2]}n=s[e.toLowerCase()]}return n===t?null:n},overrideMimeType:function(e){return E||(c.mimeType=e),this},abort:function(e){return e=e||S,o&&o.abort(e),T(0,e),this}};d.promise(x),x.success=x.done,x.error=x.fail,x.complete=m.add,x.statusCode=function(e){if(e){var t;if(E<2)for(t in e)g[t]=[g[t],e[t]];else t=e[x.status],x.always(t)}return this},c.url=((e||c.url)+"").replace(hn,"").replace(mn,ln[1]+"//"),c.dataTypes=v.trim(c.dataType||"*").toLowerCase().split(y),c.crossDomain==null&&(a=wn.exec(c.url.toLowerCase()),c.crossDomain=!(!a||a[1]===ln[1]&&a[2]===ln[2]&&(a[3]||(a[1]==="http:"?80:443))==(ln[3]||(ln[1]==="http:"?80:443)))),c.data&&c.processData&&typeof c.data!="string"&&(c.data=v.param(c.data,c.traditional)),kn(Sn,c,n,x);if(E===2)return x;f=c.global,c.type=c.type.toUpperCase(),c.hasContent=!vn.test(c.type),f&&v.active++===0&&v.event.trigger("ajaxStart");if(!c.hasContent){c.data&&(c.url+=(gn.test(c.url)?"&":"?")+c.data,delete c.data),r=c.url;if(c.cache===!1){var N=v.now(),C=c.url.replace(bn,"$1_="+N);c.url=C+(C===c.url?(gn.test(c.url)?"&":"?")+"_="+N:"")}}(c.data&&c.hasContent&&c.contentType!==!1||n.contentType)&&x.setRequestHeader("Content-Type",c.contentType),c.ifModified&&(r=r||c.url,v.lastModified[r]&&x.setRequestHeader("If-Modified-Since",v.lastModified[r]),v.etag[r]&&x.setRequestHeader("If-None-Match",v.etag[r])),x.setRequestHeader("Accept",c.dataTypes[0]&&c.accepts[c.dataTypes[0]]?c.accepts[c.dataTypes[0]]+(c.dataTypes[0]!=="*"?", "+Tn+"; q=0.01":""):c.accepts["*"]);for(l in c.headers)x.setRequestHeader(l,c.headers[l]);if(!c.beforeSend||c.beforeSend.call(h,x,c)!==!1&&E!==2){S="abort";for(l in{success:1,error:1,complete:1})x[l](c[l]);o=kn(xn,c,n,x);if(!o)T(-1,"No Transport");else{x.readyState=1,f&&p.trigger("ajaxSend",[x,c]),c.async&&c.timeout>0&&(u=setTimeout(function(){x.abort("timeout")},c.timeout));try{E=1,o.send(b,T)}catch(k){if(!(E<2))throw k;T(-1,k)}}return x}return x.abort()},active:0,lastModified:{},etag:{}});var Mn=[],_n=/\?/,Dn=/(=)\?(?=&|$)|\?\?/,Pn=v.now();v.ajaxSetup({jsonp:"callback",jsonpCallback:function(){var e=Mn.pop()||v.expando+"_"+Pn++;return this[e]=!0,e}}),v.ajaxPrefilter("json jsonp",function(n,r,i){var s,o,u,a=n.data,f=n.url,l=n.jsonp!==!1,c=l&&Dn.test(f),h=l&&!c&&typeof a=="string"&&!(n.contentType||"").indexOf("application/x-www-form-urlencoded")&&Dn.test(a);if(n.dataTypes[0]==="jsonp"||c||h)return s=n.jsonpCallback=v.isFunction(n.jsonpCallback)?n.jsonpCallback():n.jsonpCallback,o=e[s],c?n.url=f.replace(Dn,"$1"+s):h?n.data=a.replace(Dn,"$1"+s):l&&(n.url+=(_n.test(f)?"&":"?")+n.jsonp+"="+s),n.converters["script json"]=function(){return u||v.error(s+" was not called"),u[0]},n.dataTypes[0]="json",e[s]=function(){u=arguments},i.always(function(){e[s]=o,n[s]&&(n.jsonpCallback=r.jsonpCallback,Mn.push(s)),u&&v.isFunction(o)&&o(u[0]),u=o=t}),"script"}),v.ajaxSetup({accepts:{script:"text/javascript, application/javascript, application/ecmascript, application/x-ecmascript"},contents:{script:/javascript|ecmascript/},converters:{"text script":function(e){return v.globalEval(e),e}}}),v.ajaxPrefilter("script",function(e){e.cache===t&&(e.cache=!1),e.crossDomain&&(e.type="GET",e.global=!1)}),v.ajaxTransport("script",function(e){if(e.crossDomain){var n,r=i.head||i.getElementsByTagName("head")[0]||i.documentElement;return{send:function(s,o){n=i.createElement("script"),n.async="async",e.scriptCharset&&(n.charset=e.scriptCharset),n.src=e.url,n.onload=n.onreadystatechange=function(e,i){if(i||!n.readyState||/loaded|complete/.test(n.readyState))n.onload=n.onreadystatechange=null,r&&n.parentNode&&r.removeChild(n),n=t,i||o(200,"success")},r.insertBefore(n,r.firstChild)},abort:function(){n&&n.onload(0,1)}}}});var Hn,Bn=e.ActiveXObject?function(){for(var e in Hn)Hn[e](0,1)}:!1,jn=0;v.ajaxSettings.xhr=e.ActiveXObject?function(){return!this.isLocal&&Fn()||In()}:Fn,function(e){v.extend(v.support,{ajax:!!e,cors:!!e&&"withCredentials"in e})}(v.ajaxSettings.xhr()),v.support.ajax&&v.ajaxTransport(function(n){if(!n.crossDomain||v.support.cors){var r;return{send:function(i,s){var o,u,a=n.xhr();n.username?a.open(n.type,n.url,n.async,n.username,n.password):a.open(n.type,n.url,n.async);if(n.xhrFields)for(u in n.xhrFields)a[u]=n.xhrFields[u];n.mimeType&&a.overrideMimeType&&a.overrideMimeType(n.mimeType),!n.crossDomain&&!i["X-Requested-With"]&&(i["X-Requested-With"]="XMLHttpRequest");try{for(u in i)a.setRequestHeader(u,i[u])}catch(f){}a.send(n.hasContent&&n.data||null),r=function(e,i){var u,f,l,c,h;try{if(r&&(i||a.readyState===4)){r=t,o&&(a.onreadystatechange=v.noop,Bn&&delete Hn[o]);if(i)a.readyState!==4&&a.abort();else{u=a.status,l=a.getAllResponseHeaders(),c={},h=a.responseXML,h&&h.documentElement&&(c.xml=h);try{c.text=a.responseText}catch(p){}try{f=a.statusText}catch(p){f=""}!u&&n.isLocal&&!n.crossDomain?u=c.text?200:404:u===1223&&(u=204)}}}catch(d){i||s(-1,d)}c&&s(u,f,c,l)},n.async?a.readyState===4?setTimeout(r,0):(o=++jn,Bn&&(Hn||(Hn={},v(e).unload(Bn)),Hn[o]=r),a.onreadystatechange=r):r()},abort:function(){r&&r(0,1)}}}});var qn,Rn,Un=/^(?:toggle|show|hide)$/,zn=new RegExp("^(?:([-+])=|)("+m+")([a-z%]*)$","i"),Wn=/queueHooks$/,Xn=[Gn],Vn={"*":[function(e,t){var n,r,i=this.createTween(e,t),s=zn.exec(t),o=i.cur(),u=+o||0,a=1,f=20;if(s){n=+s[2],r=s[3]||(v.cssNumber[e]?"":"px");if(r!=="px"&&u){u=v.css(i.elem,e,!0)||n||1;do a=a||".5",u/=a,v.style(i.elem,e,u+r);while(a!==(a=i.cur()/o)&&a!==1&&--f)}i.unit=r,i.start=u,i.end=s[1]?u+(s[1]+1)*n:n}return i}]};v.Animation=v.extend(Kn,{tweener:function(e,t){v.isFunction(e)?(t=e,e=["*"]):e=e.split(" ");var n,r=0,i=e.length;for(;r<i;r++)n=e[r],Vn[n]=Vn[n]||[],Vn[n].unshift(t)},prefilter:function(e,t){t?Xn.unshift(e):Xn.push(e)}}),v.Tween=Yn,Yn.prototype={constructor:Yn,init:function(e,t,n,r,i,s){this.elem=e,this.prop=n,this.easing=i||"swing",this.options=t,this.start=this.now=this.cur(),this.end=r,this.unit=s||(v.cssNumber[n]?"":"px")},cur:function(){var e=Yn.propHooks[this.prop];return e&&e.get?e.get(this):Yn.propHooks._default.get(this)},run:function(e){var t,n=Yn.propHooks[this.prop];return this.options.duration?this.pos=t=v.easing[this.easing](e,this.options.duration*e,0,1,this.options.duration):this.pos=t=e,this.now=(this.end-this.start)*t+this.start,this.options.step&&this.options.step.call(this.elem,this.now,this),n&&n.set?n.set(this):Yn.propHooks._default.set(this),this}},Yn.prototype.init.prototype=Yn.prototype,Yn.propHooks={_default:{get:function(e){var t;return e.elem[e.prop]==null||!!e.elem.style&&e.elem.style[e.prop]!=null?(t=v.css(e.elem,e.prop,!1,""),!t||t==="auto"?0:t):e.elem[e.prop]},set:function(e){v.fx.step[e.prop]?v.fx.step[e.prop](e):e.elem.style&&(e.elem.style[v.cssProps[e.prop]]!=null||v.cssHooks[e.prop])?v.style(e.elem,e.prop,e.now+e.unit):e.elem[e.prop]=e.now}}},Yn.propHooks.scrollTop=Yn.propHooks.scrollLeft={set:function(e){e.elem.nodeType&&e.elem.parentNode&&(e.elem[e.prop]=e.now)}},v.each(["toggle","show","hide"],function(e,t){var n=v.fn[t];v.fn[t]=function(r,i,s){return r==null||typeof r=="boolean"||!e&&v.isFunction(r)&&v.isFunction(i)?n.apply(this,arguments):this.animate(Zn(t,!0),r,i,s)}}),v.fn.extend({fadeTo:function(e,t,n,r){return this.filter(Gt).css("opacity",0).show().end().animate({opacity:t},e,n,r)},animate:function(e,t,n,r){var i=v.isEmptyObject(e),s=v.speed(t,n,r),o=function(){var t=Kn(this,v.extend({},e),s);i&&t.stop(!0)};return i||s.queue===!1?this.each(o):this.queue(s.queue,o)},stop:function(e,n,r){var i=function(e){var t=e.stop;delete e.stop,t(r)};return typeof e!="string"&&(r=n,n=e,e=t),n&&e!==!1&&this.queue(e||"fx",[]),this.each(function(){var t=!0,n=e!=null&&e+"queueHooks",s=v.timers,o=v._data(this);if(n)o[n]&&o[n].stop&&i(o[n]);else for(n in o)o[n]&&o[n].stop&&Wn.test(n)&&i(o[n]);for(n=s.length;n--;)s[n].elem===this&&(e==null||s[n].queue===e)&&(s[n].anim.stop(r),t=!1,s.splice(n,1));(t||!r)&&v.dequeue(this,e)})}}),v.each({slideDown:Zn("show"),slideUp:Zn("hide"),slideToggle:Zn("toggle"),fadeIn:{opacity:"show"},fadeOut:{opacity:"hide"},fadeToggle:{opacity:"toggle"}},function(e,t){v.fn[e]=function(e,n,r){return this.animate(t,e,n,r)}}),v.speed=function(e,t,n){var r=e&&typeof e=="object"?v.extend({},e):{complete:n||!n&&t||v.isFunction(e)&&e,duration:e,easing:n&&t||t&&!v.isFunction(t)&&t};r.duration=v.fx.off?0:typeof r.duration=="number"?r.duration:r.duration in v.fx.speeds?v.fx.speeds[r.duration]:v.fx.speeds._default;if(r.queue==null||r.queue===!0)r.queue="fx";return r.old=r.complete,r.complete=function(){v.isFunction(r.old)&&r.old.call(this),r.queue&&v.dequeue(this,r.queue)},r},v.easing={linear:function(e){return e},swing:function(e){return.5-Math.cos(e*Math.PI)/2}},v.timers=[],v.fx=Yn.prototype.init,v.fx.tick=function(){var e,n=v.timers,r=0;qn=v.now();for(;r<n.length;r++)e=n[r],!e()&&n[r]===e&&n.splice(r--,1);n.length||v.fx.stop(),qn=t},v.fx.timer=function(e){e()&&v.timers.push(e)&&!Rn&&(Rn=setInterval(v.fx.tick,v.fx.interval))},v.fx.interval=13,v.fx.stop=function(){clearInterval(Rn),Rn=null},v.fx.speeds={slow:600,fast:200,_default:400},v.fx.step={},v.expr&&v.expr.filters&&(v.expr.filters.animated=function(e){return v.grep(v.timers,function(t){return e===t.elem}).length});var er=/^(?:body|html)$/i;v.fn.offset=function(e){if(arguments.length)return e===t?this:this.each(function(t){v.offset.setOffset(this,e,t)});var n,r,i,s,o,u,a,f={top:0,left:0},l=this[0],c=l&&l.ownerDocument;if(!c)return;return(r=c.body)===l?v.offset.bodyOffset(l):(n=c.documentElement,v.contains(n,l)?(typeof l.getBoundingClientRect!="undefined"&&(f=l.getBoundingClientRect()),i=tr(c),s=n.clientTop||r.clientTop||0,o=n.clientLeft||r.clientLeft||0,u=i.pageYOffset||n.scrollTop,a=i.pageXOffset||n.scrollLeft,{top:f.top+u-s,left:f.left+a-o}):f)},v.offset={bodyOffset:function(e){var t=e.offsetTop,n=e.offsetLeft;return v.support.doesNotIncludeMarginInBodyOffset&&(t+=parseFloat(v.css(e,"marginTop"))||0,n+=parseFloat(v.css(e,"marginLeft"))||0),{top:t,left:n}},setOffset:function(e,t,n){var r=v.css(e,"position");r==="static"&&(e.style.position="relative");var i=v(e),s=i.offset(),o=v.css(e,"top"),u=v.css(e,"left"),a=(r==="absolute"||r==="fixed")&&v.inArray("auto",[o,u])>-1,f={},l={},c,h;a?(l=i.position(),c=l.top,h=l.left):(c=parseFloat(o)||0,h=parseFloat(u)||0),v.isFunction(t)&&(t=t.call(e,n,s)),t.top!=null&&(f.top=t.top-s.top+c),t.left!=null&&(f.left=t.left-s.left+h),"using"in t?t.using.call(e,f):i.css(f)}},v.fn.extend({position:function(){if(!this[0])return;var e=this[0],t=this.offsetParent(),n=this.offset(),r=er.test(t[0].nodeName)?{top:0,left:0}:t.offset();return n.top-=parseFloat(v.css(e,"marginTop"))||0,n.left-=parseFloat(v.css(e,"marginLeft"))||0,r.top+=parseFloat(v.css(t[0],"borderTopWidth"))||0,r.left+=parseFloat(v.css(t[0],"borderLeftWidth"))||0,{top:n.top-r.top,left:n.left-r.left}},offsetParent:function(){return this.map(function(){var e=this.offsetParent||i.body;while(e&&!er.test(e.nodeName)&&v.css(e,"position")==="static")e=e.offsetParent;return e||i.body})}}),v.each({scrollLeft:"pageXOffset",scrollTop:"pageYOffset"},function(e,n){var r=/Y/.test(n);v.fn[e]=function(i){return v.access(this,function(e,i,s){var o=tr(e);if(s===t)return o?n in o?o[n]:o.document.documentElement[i]:e[i];o?o.scrollTo(r?v(o).scrollLeft():s,r?s:v(o).scrollTop()):e[i]=s},e,i,arguments.length,null)}}),v.each({Height:"height",Width:"width"},function(e,n){v.each({padding:"inner"+e,content:n,"":"outer"+e},function(r,i){v.fn[i]=function(i,s){var o=arguments.length&&(r||typeof i!="boolean"),u=r||(i===!0||s===!0?"margin":"border");return v.access(this,function(n,r,i){var s;return v.isWindow(n)?n.document.documentElement["client"+e]:n.nodeType===9?(s=n.documentElement,Math.max(n.body["scroll"+e],s["scroll"+e],n.body["offset"+e],s["offset"+e],s["client"+e])):i===t?v.css(n,r,i,u):v.style(n,r,i,u)},n,o?i:t,o,null)}})}),e.jQuery=e.$=v,typeof define=="function"&&define.amd&&define.amd.jQuery&&define("jquery",[],function(){return v})})(window);
 
</script>
 
</script>
−
<script src="https://2014hs.igem.org/Mingdao_JS_museutils?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
−
  <script src="https://2014hs.igem.org/Mingdao_JS_jquery.tobrowserwidth?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
−
  <script src="https://2014hs.igem.org/Mingdao_JS_jquery.musemenu?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
−
  <script src="https://2014hs.igem.org/Mingdao_JS_jquery.watch?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
−
  <script src="https://2014hs.igem.org/Mingdao_JS_jquery.scrolleffects?action=raw&ctype=text/javascript" type="text/javascript"></script>
 
−
  <!-- Other scripts -->
 
 
   <script type="text/javascript">
 
   <script type="text/javascript">
−
   $(document).ready(function() { try {
+
   window.jQuery || document.write('\x3Cscript src="https://2015.igem.org/Team:Mingdao/Javascript-jquery-1.8.3?
−
Muse.Utils.transformMarkupToFixBrowserProblemsPreInit();/* body */
+
action=raw&ctype=text/javascript" type="text/javascript">\x3C/script>');
−
$('.browser_width').toBrowserWidth();/* browser width elements */
+
−
Muse.Utils.prepHyperlinks(true);/* body */
+
−
Muse.Utils.initWidget('.MenuBar', function(elem) { return $(elem).museMenu(); });/* unifiedNavBar */
+
−
$('#u2764').registerPositionScrollEffect([{"speed":[0,1],"in":[-Infinity,36.95]},{"speed":[0,1],"in":[36.95,Infinity]}]);/* scroll effect */
+
−
Muse.Utils.fullPage('#page');/* 100% height page */
+
−
Muse.Utils.showWidgetsWhenReady();/* body */
+
−
Muse.Utils.transformMarkupToFixBrowserProblems();/* body */
+
−
} catch(e) { Muse.Assert.fail('Error calling selector function:' + e); }});
+
 
</script>
 
</script>
 +
  <script src="https://2015.igem.org/Team:Mingdao/Javascript-museutils?
 +
action=raw&amp;ctype=text/javascript" type="text/javascript"></script>
 +
  <script src="https://2015.igem.org/Team:Mingdao/Javascript-jquery.watch?
 +
action=raw&amp;ctype=text/javascript" type="text/javascript"></script>
 +
  <script src="https://2015.igem.org/Team:Mingdao/Javascript-jquery.musemenu?
 +
action=raw&amp;ctype=text/javascript" type="text/javascript"></script>
 +
  <!-- Other scripts -->
 +
  <script>if (window.runOnloadHook) runOnloadHook();</script>
 
<!-- JS includes -->
 
<!-- JS includes -->
 
   <script type="text/javascript">
 
   <script type="text/javascript">
Line 455: Line 2,480:
 
Muse.Utils.showWidgetsWhenReady();/* body */
 
Muse.Utils.showWidgetsWhenReady();/* body */
 
Muse.Utils.transformMarkupToFixBrowserProblems();/* body */
 
Muse.Utils.transformMarkupToFixBrowserProblems();/* body */
−
} catch(e) { Muse.Assert.fail('Error calling selector function:' + e); }});
+
} catch(e) { }});
 
</script>
 
</script>
−
  </body>
+
 
 +
</body>
 
</html>
 
</html>

Latest revision as of 15:28, 18 September 2015

Parts

Gene Block

*

*

In our project, we cloned two genes, SR and SRPK from the materials kindly gave by Dr. Woan-Yuh Tarn at Academia Sinica in Taiwan. Phosphoserine and arginine (highest nitrogen contents among 20 amino acids) play important roles in fire retardance. We found a novel fire retardant protein, SR protein, through protein database mining. SR proteins are post-translationally phosphorylated by SR protein kinase (SRPK). We co-transformed E. coli BL21 with two vectors simultaneously to express phosphorylated SR proteins.

Gene

Serine/arginine-rich splicing factor 1 (SRSF1) is a protein found on chromosome 17 in humans. SRSF1 is also known as alternative splicing factor 1 (ASF1), pre-mRNA-splicing factor SF2 (SF2), which is involved in alternative splicing, mRNA nuclear export and translation. It contains two functional domains. One in the N-terminus is RNA recognition motifs which interact with RNA and splicing factors. The other in the C-terminus is rich in serine and arginine residues (SR domain) and regulate the protein activity through phosphorylation by SR protein kinase.

 

SRSF protein kinase 1 (SRPK1) is a serine/arginine-rich protein specific kinase, which phosphorylates SR proteins on serine residues in SR domain. It plays an important role in the regulation of SR protein activity. Disorders in SRPK1 may be observed in certain cancer cells.

 

BioBrick

We cloned the genes of SR and SRPK as well as the composite DNA fragments of Ptac-GST-SR and Pt7-SRPK-His onto the standard backbone of pSB1C3 biobrick.

SR/pSB1C3

Part No.: BBa_K1608000

We designed two primers to amplify the SR genes by PCR, followed by XbaI and PstI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation.

Primers:

1. SR-XbaI-F: 5'- AAAAGGTCTAGATGTCGGGAGGTGGTGTGATTCGTG -3'

2. SR-SpeI-PstI-R: 5'-AAGAAACTGCAGCGGCCGCTACTAGTATTATGTACGAGAGCGAGA -3'

PCR product size: ~785 bp

 

SRPK/pSB1C3

Part No.: BBa_K1608001

We designed two primers to amplify SRPK1 gene by PCR, followed by EcoRI and SpeI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. However, there’re two PstI sites on the sequence of SRPK1 gene. We’re trying to mutate them to fit the standard biobrick parts.

Primers:

1. SRPK1-EcoRI-XbaI-F: 5'- AAAAGAATTCGCGGCCGCTTCTAGATGGAGCGGAAAGTGCTTGCGCTCCAG -3'

2. SRPK1-SpeI-R: 5'- GGGGGGACTAGTATTAGGAGTTAAGCCAAGGGTG -3'

PCR product size: ~1995 bp

Plasmid map:

 

In order to express SR and SRPK gene more conveniently, we made two composite biobricks with promoter, RBS and a fusion tag.

GST-SR/pSB1C3

Part No.: BBa_K1608002

We designed two primers to amplify Ptac-GST-SR DNA fragment by PCR from SR/pGEX-2T, followed by XbaI and PstI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. The SR gene is fused with GST tag and driven by Ptac promoter along with a lac operator downstream.

Primers:

1. Ptac (pGEX)-XbaI-F: 5'- GAGGGGTCTAGAGTTTTTGCGCCGACATCATAAC -3'

2. SR-SpeI-PstI-R: 5'- AAGAAACTGCAGCGGCCGCTACTAGTATTATGTACGAGAGCGAGA -3'

PCR product size: ~1572 bp

Plasmid map:

 

In order to express SR and SRPK gene more conveniently, we made two composite biobricks with promoter, RBS and a fusion tag.

SRPK-His/pSB1C3

Part No.: BBa_K1608003

 We designed two primers to amplify Pt7-SRPK1-His DNA fragment by PCR from SRPK/pET-29b, followed by EcoRI and SpeI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. However, there’re two PstI sites one XbaI site on the amplified sequence. We’re trying to mutate them to fit the standard biobrick parts. The SRPK1 gene is fused with 6XHis tag and driven by Pt7 promoter along with a lac operator downstream.

Primers:

1. Pt7(pET)-EcoRI-XbaI-F: 5'- AAAAAAGAATTCGCGGCCGCTTCTAGAGCACGATGCGTCCGGCGTA -3'

2. His (pET)-SpeI-R: 5'- AGGGAAACTAGTATCAGTGGTGGTGGTGGTGGT -3'

PCR product size: ~2294 bp

Plasmid map:

Data

(1) PCR of SR, SRPK, Ptac-GST-SR and Pt7-SRPK-His

Lanes:

1. 1kb DNA marker

2. Pt7-SRPK-His: 2294 bp

3. Ptac-GST-SR: 1572 bp

4. SR: 785 bp

5. SRPK: 1995 bp

(2) Restriction enzyme digestion of inserts and vectors

Lanes:

1. 1kb DNA marker

2. Pt7-SRPK-His: EcoRI + SpeI → 2294 bp

3. SRPK: EcoRI + SpeI →1995 bp

4. pSB1C3: EcoRI + SpeI → 2047 + 1092 bp*

5. Ptac-GST-SR: XbaI + PstI→ 1572 bp

6. SR: XbaI + PstI → 785 bp

7. pSB1C3: XbaI + PstI → 2044 + 1095 bp*

 

*Bands shifted were caused by the fluorescent DNA loading dye. We confirmed with the specialist and sequenced the plasmid. The plasmids were correct but it bothers us for a long time. Just be very careful to use a ready-to-use fluorescent DNA loading dye.

(3)Colony PCR for SRPK/pSB1C3 and SR/pSB1C3

Left panel: Colony PCR with primers of SRPK1-EcoRI-XbaI-F and VR to check SRPK/pSB1C3 (2162 bp)

 

Right panel: Colony PCR with primers of SR-XbaI-F + VR to check SR/pSB1C3 (941 bp)

 

(4)Colony PCR for Ptac-GST-SR/pSB1C3 and Pt7-SRPK-His/pSB1C3

Lane 1 & 17: 1kb DNA marker

Lane 2-8: Colony PCR with primers of Ptac (pGEX)-XbaI-F and VR to check Ptac-GST-SR/pSB1C3 (1731 bp)

Lane 9-16: Colony PCR with primers of Pt7(pET)-EcoRI-XbaI-F and VR to check Pt7-SRPK-His/pSB1C3 (2452 bp)

 

(5) Restriction enzyme check for SRPK/pSB1C3, SR/pSB1C3 and Pt7-SRPK-His/pSB1C3

Left panel:

1. SRPK/pSB1C3: EcoRI → 4036 bp

2. SRPK/pSB1C3: PstI → 2747 + 1238 + 51 bp

3. 1kb DNA marker

4. SR/pSB1C3: XbaI → 2815 bp

5. SR/pSB1C3: EcoRI + PstI → 2029 + 786 bp

 

Right panel:

1. Pt7-SRPK-His/pSB1C3: EcoRI → 4326 bp

2. Pt7-SRPK-His/pSB1C3: PstI → 2980 + 1295 + 51 bp

3. 1kb DNA marker

(6) Restriction enzyme check for Ptac-GST-SR/pSB1C3

Lanes:

1. 1kb DNA marker

2. Ptac-GST-SR/pSB1C3: uncut

3. Ptac-GST-SR/pSB1C3: BamHI → 3614 bp

4. Ptac-GST-SR/pSB1C3: EcoRI + PstI → 1581 + 2033 bp

(7) Sequencing data with primers of VF2 and VR

a. SR/pSB1C3 (VF2 & VR)

b. SRPK/pSB1C3 (VF2 & VR)

c. Ptac-GST-SR/pSB1C3 (VF2 & VR)

d. Pt7-SRPK-His/pSB1C3 (VF2 & VR)

 

Download

 

Vector

In order to express proteins abundantly and efficiently, we transferred the genes of SR and SRPK into two commercial expression vectors of pGEX-2T and pET-29b, respectively.

SR/pGEX-2T

We cloned SR gene with BamHI and XbaI onto pGEX-2T, in which the gene expression is fused with GST tag in the 5’ end and driven under Ptac promoter as well as regulated by lac operator. The pGEX-2T backbone contains lacI repressor gene, pBR322 replication origin and ampicillin resistance cassette (AmpR).

Plasmid map:

 

SRPK/pET-29b

We cloned SRPK gene with EcoRV and HindIII onto pET-29b, in which the gene expression is fused with 6XHis tag in the 3’ end and driven under Pt7 promoter as well as regulated by lac operator. The pET-29b backbone contains lacI repressor gene, ColE1 replication origin and kanamycin resistance cassette (KanR).

Plasmid map:

DATA

 

Protein Induction

We co-transformed SR/pGEX-2T and SRPK/pET-29b into E. coli BL21, which expresses T7 RNA polymerase under LacI repressor regulation. The bacteria were cultured in LB media supplemented with ampicillin and kanamycin. IPTG was added when the bacteria grew to OD600 value at 0.6-1.0. And the protein lysates were collected after 4 hours of IPTG induction. SDS-PAGE, Coomassie Blue Staining and Western Blotting were performed through the manufacturers’ instruction.

Coomassie Blue Staining

Lanes of lysates:

1. Control: pGEX-2T & pET-29b

2. SR: SR/pGEX-2T & pET-29b

3. SRPK: pGEX-2T & SRPK1/pET-29b

 

Result:

SR was expressed at the expected size of 53 kDa. and SRPK was expressed at the expected size of 93 kDa. In addition, free forms of GST were observed at 27 kDa in the lanes 1 & 3.

 

Western Blotting

(1) SRPK-His protein expression

Procedure: ↓ mouse α-His primary Ab ↓ goat α-mouse IgG-AP secondary Ab ↓ BCIP/NBT chromogen Lanes of lysates: 1. Control: pGEX-2T & pET-29b 2. SR: SR/pGEX-2T & pET-29b 3. SRPK: pGEX-2T & SRPK1/pET-29b 4. P-SR: SR/pGEX-2T & SRPK1/pET-29b Result: SRPK-His was detected by anti-His antibody at the expected size of 93 kDa, as shown in the lanes 3 & 4.

(2) GST-SR protein expression

Procedure: ↓ rabbit α-GST primary Ab ↓ goat α-rabbit IgG-HRP secondary Ab ↓ DAB chromogen Lanes of lysates: 1. Control: pGEX-2T & pET-29b 2. SR: SR/pGEX-2T & pET-29b 3. SRPK: pGEX-2T & SRPK1/pET-29b 4. P-SR: SR/pGEX-2T & SRPK1/pET-29b

Result

GST-SR was detected by anti-GST antibody at the expected size of 53 kDa, as shown in the lanes 2 & 4. It is worth to note that the bands in lane 4 were shifted around 53 kDa, indicating protein modification of GST-SR possibly by SRPK kinase phosphorylation (SR and SRPK exist in the bacteria all together). Moreover, the free forms of GST around 27 kDa were also observed.

 

Summary

We cloned the genes encoding SR and SRPK to BioBricks (pSB1C3) and expression vectors (pGEX-2T and pET-29b). After protein induction by IPTG in E. coli BL21, the SR and SRPK as well as phosphorylated SR proteins were detected in SDS-PAGE with Coomassie Blue staining and Western Blotting by anti-His antibody and anti-GST antibody. The results showed that we successfully expressed phosphorylated SR proteins in E. coli bacteria by transformed with SR/pGEX-2T and SRPK/pET-29b.