Difference between revisions of "Team:EPF Lausanne/Part Collection"
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<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>PAM rich URS J23117</h2> | + | <h2>PAM rich URS J23117 promoter</h2> |
<p>This promoter is composed of the weak J23117 promoter, BBa_J23117, fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind as dCas9 cannot bind without a PAM sequence.</p> | <p>This promoter is composed of the weak J23117 promoter, BBa_J23117, fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind as dCas9 cannot bind without a PAM sequence.</p> | ||
</div> | </div> | ||
| Line 65: | Line 65: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA Z0</h2> | + | <h2>sgRNA Z0 expressing cassette</h2> |
<p>the sgRNA (single guide-RNA) Z0 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA (single guide-RNA) Z0 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
| Line 83: | Line 83: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA Z4</h2> | + | <h2>sgRNA Z4 expressing cassette</h2> |
<p>the sgRNA Z4 activates PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA Z4 activates PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
| Line 101: | Line 101: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA Z35</h2> | + | <h2>sgRNA Z35 expressing cassette</h2> |
<p>the sgRNA Z35 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA Z35 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
| Line 118: | Line 118: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>PAM rich URS J23117Alt</h2> | + | <h2>PAM rich URS J23117Alt promoter</h2> |
<p>This promoter was made on the same template as PAM rich URS J23117 promoter, but was mutated to obtain different sequences for sgRNA targeting. It is also fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind, as dCas9 cannot bind without a PAM sequence.</p> | <p>This promoter was made on the same template as PAM rich URS J23117 promoter, but was mutated to obtain different sequences for sgRNA targeting. It is also fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind, as dCas9 cannot bind without a PAM sequence.</p> | ||
</div> | </div> | ||
| Line 136: | Line 136: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA X0</h2> | + | <h2>sgRNA X0 expressing cassette</h2> |
<p>the sgRNA (single guide-RNA) X0 inhibit PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA (single guide-RNA) X0 inhibit PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
| Line 154: | Line 154: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA X4</h2> | + | <h2>sgRNA X4 expressing cassette</h2> |
<p>the sgRNA X4 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA X4 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
| Line 172: | Line 172: | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
| − | <h2>sgRNA X35</h2> | + | <h2>sgRNA X35 expressing cassette</h2> |
<p>the sgRNA X35 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | <p>the sgRNA X35 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.</p> | ||
</div> | </div> | ||
Revision as of 19:42, 16 September 2015
E. coli
dCas9-ω
dCa9s-ω is our main tool to build our project in E. coli. This catalytically dead Cas9 fused to the RNA polymerase ω subunit can act as an activator or as an inhibitor when provided the right guide RNA.
PAM rich URS J23117 promoter
This promoter is composed of the weak J23117 promoter, BBa_J23117, fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind as dCas9 cannot bind without a PAM sequence.
sgRNA Z0 expressing cassette
the sgRNA (single guide-RNA) Z0 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA Z4 expressing cassette
the sgRNA Z4 activates PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA Z35 expressing cassette
the sgRNA Z35 inhibit PAM rich URS J23117 promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence Z35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
PAM rich URS J23117Alt promoter
This promoter was made on the same template as PAM rich URS J23117 promoter, but was mutated to obtain different sequences for sgRNA targeting. It is also fused to a PAM rich upstream regulatory region (URS) that is needed for dCas9 to bind, as dCas9 cannot bind without a PAM sequence.
sgRNA X0 expressing cassette
the sgRNA (single guide-RNA) X0 inhibit PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X0 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA X4 expressing cassette
the sgRNA X4 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X4 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
sgRNA X35 expressing cassette
the sgRNA X35 activates PAM rich URS J23117Alt promoter when in complex with dCas9-ω. This part is an expressing cassette under the constitutive promoter pBAD. The sequence X35 makes this part unique, and the scaffold is a common part to all the sgRNAs and is recognized by dCas9-ω.
S. cerevisiae
