Team:Evry Paris-Saclay/Part Collection


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Genopole Campus 1, Batiment 6, 91030 Evry Cedex, France
+33 7 69 96 68 31
igemevry@gmail.com

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Design & Developpement by
IGEM EVRY GENOPOLE

COMPOSITE PARTS

In our project we tried to repurpose a peptide communication machinery from a phage of the Gram-positive Bacillus subtilis to the cells of Gram-negative Escherichia coli, and quickly discovered that crossing the inter-species barrier in bioengineering is a challenging task. This Part Collection consists of a series of promoter variants designed to debug the mechanism of transcriptional regulation in the pAimX promoter that is key to the signalling system. The Collection helped us successfully identify an internal terminator, investigate a potential anti-termination mechanism, and build a hybrid promoter that converts the native transcriptional activator (AimR) into a transcriptional repressor.

Part Number Part Name
BBa_K2675020 pAimX(full) promoter of phage phi3T
BBa_K2675021 pAimX(short) promoter of phage phi3T
BBa_K2675022 pAimX(short)-J23110 hybrid promoter
BBa_K2675023 pAimX(full-1)-J23110 hybrid promoter
BBa_K2675024 pAimX(full-2)-J23110 hybrid promoter
BBa_K2675025 pAimX(full-3)-J23110 hybrid promoter
BBa_K2675090 pAimX(full-noTerminator-v1) promoter of phage phi3T
BBa_K2675091 pAimX(full-noTerminator-v2) promoter of phage phi3T
BBa_K2675092 pAimX(full-noTerminator-v3) promoter of phage phi3T
BBa_K2675093 pAimX(full-noTerminator-v4) promoter of phage phi3T
BBa_K2675094 pAimX(full-noTerminator-v5) promoter of phage phi3T
BBa_K2675095 pAimX(full-noTerminator-v6) promoter of phage phi3T
BBa_K2675096 pAimX(full-noTerminator-v7) promoter of phage phi3T
BBa_K2675097 pAimX(full-noTerminator-v8) promoter of phage phi3T

When placed upstream of a reporter gene, promoters from this Part Collection showed variable transcription strengths and degrees of regulation that provided valuable insights for our project and facilitated the porting of the peptide communication machinery from B. subtilis to E. coli.

To build this Part Collection the BBa_K2675080 and BBa_K2675081 were a key tool.

Part Number Part Name
BBa_K2675050 sfGFP-LVAtag expression cassette under the control of pAimX(full) promoter of phage phi3T
BBa_K2675051 sfGFP-LVAtag expression cassette under the control of pAimX(short) promoter of phage phi3T
BBa_K2675052 sfGFP-LVAtag expression cassette under the control of pAimX(short)-J23110 hybrid promoter
BBa_K2675053 sfGFP-LVAtag expression cassette under the control of pAimX(full-1)-J23110 hybrid promoter
BBa_K2675054 sfGFP-LVAtag expression cassette under the control of pAimX(full-2)-J23110 hybrid promoter
BBa_K2675055 sfGFP-LVAtag expression cassette under the control of pAimX(full-3)-J23110 hybrid promoter
BBa_K2675057 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full) promoter of phage phi3T
BBa_K2675058 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(short) promoter of phage phi3T
BBa_K2675059 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(short) promoter of phage phi3T
BBa_K2675190 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v1) promoter of phage phi3T
BBa_K2675191 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v2) promoter of phage phi3T
BBa_K2675192 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v3) promoter of phage phi3T
BBa_K2675193 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v4) promoter of phage phi3T
BBa_K2675194 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v5) promoter of phage phi3T
BBa_K2675195 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v6) promoter of phage phi3T
BBa_K2675196 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v7) promoter of phage phi3T
BBa_K2675197 sfGFP-LVAtag expression cassette under the control of J23110 and pAimX(full-noTerminator-v8) promoter of phage phi3T
BBa_K2675060 sfGFP expression cassette under the control of pAimX(full) promoter of phage phi3T
BBa_K2675061 sfGFP expression cassette under the control of pAimX(short) promoter of phage phi3T
BBa_K2675062 sfGFP expression cassette under the control of pAimX(short)-J23110 hybrid promoter
BBa_K2675063 sfGFP expression cassette under the control of pAimX(full-1)-J23110 hybrid promoter
BBa_K2675064 sfGFP expression cassette under the control of pAimX(full-2)-J23110 hybrid promoter
BBa_K2675065 sfGFP expression cassette under the control of pAimX(full-3)-J23110 hybrid promoter