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Team:EPF-Lausanne/Parts
From 2008.igem.org
Cloning Scheme
First plasmid : the basic network components
Second plasmid : the testing component
Parts submitted to the registry
- TetR coding sequence with RBS and terminator :
- Coding sequence of the Tetracycline repressor with the ribosome binding site and a double terminator. The tetR comes from transposon tn10 and has a LVA tail (SsrA) [cf. C0040 part].
- This part has been successfully sequenced.
- Constitutive RhlR Receiver
- This part has been successfully sequenced.
- RhlI with RBS and terminator
- RFP controlled by a TetR promoter :
- Coding sequence of the RFP with a ribosome binding site and p(tetR) as promoter. Thus, the transcription of RFP is inhibited when TetR is present.
- This part has been successfully sequenced.
- New part created by 2-steps PCR. RBS (B0034) + luxI + Terminator (B0010+B0012) :
- Coding sequence for the Lux Inducer with a ribosome binding site and terminator. This biobrick is an alternative to the F1610 which didn't work
- TetR cds with RBS and Terminator + p(tetR), RFP with RBS
- The TetR sequence (with a ribosome binding site and terminator) is bound to the part containing the promoter of tetR, a ribosome binding site and the red fluorescent protein (RFP). Thus, the production of RFP directly depends on the production of TetR. The transcription of RFP is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
- This part has been successfully sequenced.
- TetR cds with RBS and Terminator + p(tetR, RFP with RBS + luxI with RBS and terminator
- The TetR sequence (with a ribosome binding site and terminator) BBa_K092000 is bound to the part containing the promoter of tetR and the red fluorescent protein (RFP) with a ribosome binding site. A luxI part (with RBS and terminator) is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the tetR promoter. The transcription of RFP and luxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
- Sequence that codes the same amino acid sequence than LacI but coded with a codon optimized sequence. It will thus have a different rate of translation than LacI.
- Source of LacIm : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
- rhlR controlled by a constitutive promoter, and lacIm with a RhlR promoter.
- BBa_K092901: Project Part
- Sequence containing : - rhlR controlled by a constitutive promoter - lacIm modified with a RhlR promoter - tetR controlled by a Lac Promoter - RFP controlled by a TetR promoter - luxI is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and luxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
Unfortunately, as we are finishing our wiki, the last ligation is running. Thus, we can not submit our project part.
