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Revision as of 02:16, 18 October 2018


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

© Copyright 2018
Design & Developpement by
IGEM EVRY GENOPOLE

BASIC PARTS

During this project, our team designed the following basic parts:


Coding sequences

Part Number Part Name
BBa_K2675000 AimR-Phi3T
BBa_K2675001 AimP-Phi3T
BBa_K2675002 OmpA-SAIRGA
BBa_K2675003 PelB-SAIRGA
BBa_K2675004 TAT-SAIRGA
BBa_K2675005 sfGFP
BBa_K2675006 sfGFP-LVAtag


Ribosomal binding sites (RBS)

Part Number Part Name
BBa_K2675010 Synthetic RBS designed for AimR of Phi3T
BBa_K2675011 Synthetic RBS designed for AimP of Phi3T
BBa_K2675012 Synthetic RBS designed for OmpA-SAIRGA
BBa_K2675013 Synthetic RBS designed for PelB-SAIRGA
BBa_K2675014 Synthetic RBS designed for TAT-SAIRGA
BBa_K2675015 Synthetic RBS designed for sfGFP
BBa_K2675016 Synthetic RBS designed for sfGFP
BBa_K2675017 Synthetic RBS designed for sfGFP


Promoters

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_K2675026 J23110 truncated promoter (-10 only) + cloning Scar
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


Terminators

Part Number Part Name
BBa_K2675030 L3S2P56 Terminator
BBa_K2675031 L3S2P21 Terminator


And a Golden Gate adapter

Part Number Part Name
BBa_K2675070 Golden Gate adapter

COMPOSITE PARTS

During this project, our team designed the following composite parts:

Part Number Part Name
BBa_K2675040 AimR-Phi3T expression cassette
BBa_K2675041 AimP-Phi3T expression cassette
BBa_K2675042 OmpA-SAIRGA expression cassette
BBa_K2675043 PelB-SAIRGA expression cassette
BBa_K2675044 TAT-SAIRGA expression cassette
BBa_K2675080 Golden Gate adapter + sfGFP
BBa_K2675081 Golden Gate adapter + sfGFP-LVAtag
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_K2675056 sfGFP-LVAtag expression cassette under the control of J23110 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 truncated promoter (-10 only) and pAimX(short) promoter of phage phi3T
BBa_K267190 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
BBa_K2675066 sfGFP expression cassette under the control of J23110 promoter

PART COLLECTION

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