Team:ETH Zurich/Parts

"What I cannot create I do not understand."
- Richard Feynmann

Overview

For completing our project, we required of many parts. Some of these parts where used from the Registry, others were created for this project. In the following section, you can see these parts.

Registry parts

Part Name

Description

Registry Number

INP-EYFP Fusion of Ice Nucleation Protein (INP) and Enhanced Yellow Fluorescent Protein (EYFP) BBa_K523013
cI cI repressor from E. coli phage lambda modified with an LVA tail for rapid degradation of the protein BBa_C0051
lacI + LVA lacI repressor from E. coli (+LVA) BBa_C0012
HylA HlyA-tag+Secretion system, allows a protein to be secreted by means of the alpha-hemolysin secretion system in E. coli BBa_K1166002
pLuxR Promoter activated by the LuxR protein complexed with the autoinducer homoserine lactone (HSL) BBa_R0062
pLldR operon lldPRD operon promoter + RBS from E. coli. In the absence of L-lactate, lldR binds to two operator sites O1 and O2 in the promoter region and inhibits expression BBa_K822000
InterLab 1 strong promoter Anderson promoter J23101 from the Anderson collection BBa_K823005 (BBa_J23101)
InterLab 2 medium-strong promoter Anderson promoter J23106 from the Anderson collection BBa_K823008 (BBa_J23106)
terminator Transcription terminator for the E.coli RNA polymerase BBa_B0012
double terminator Double terminator consisting of BBa_B0010 and BBa_B0012 BBa_B0015
very strong promoter Strong member of the family of promoters J23100 through J23119 BBa_J23100
gfp (InterLab) intermediate in screening plasmid construction containing GFP BBa_I13504
medium promoter Medium member of the family of promoters J23100 through J23119 BBa_J23118
aiiA autoinducer inactivation enzyme aiiA (no LVA, an enzyme that inactivates the acylhomoserine lactone (AHL) quorum-sensing signal BBa_C0160
LuxI autoinducer synthetase for acylhomoserine lactone (AHL), no LVA BBa_C0161
InterLab 3 weak promoter Anderson promoter J23117 from the Anderson collection BBa_K823013 (BBa_J23117)
promoter medium-weak Medium-weak member of the family of promoters J23100 through J23119 BBa_J23114
RBS RBS.3 (medium) derivative of BBa_0030 BBa_B0032
promoter plus GFP (InterLab control) J23151 inserted in the Promoter MeasKit BBa_I20270
terminator Artificial terminator, estimated %T~>90% BBa_B1006

New constructs

Part Name

Description

Registry Number

Plasmid 3 Combination of lldR operator sites O1 and O2 and promoters of variable potency. Gene expression is blocked by LldR protein (expressed separately) and induced upon addition of L-lactate. This was tested with a fusion to GFP.
PL3a lldRO1-plldR-lldRO2
PL3b J23100-lldRO1-lldRO2
PL3c J23100-lldRO1-R2oDNA-lldRO2
PL3d J23117-lldRO1-lldRO2
PL3e J23117-lldRO1-R2oDNA-lldRO2
PL3f J23118-lldRO1-lldRO2
PL3g J23118-lldRO1-R2oDNA-lldRO2
PL3h lldRO1-J23100-lldRO2
PL3i lldRO1-J23117-lldRO2
PL3j lldRO1-J23118-lldRO2
PL3k lldRO1-R2oDNA-lldRO2