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From 2008.igem.org

Cloning Scheme

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First plasmid : the basic network components

Second plasmid : the testing component

Parts submitted to the registry

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• [http://partsregistry.org/Part:BBa_K092000 BBa_K092000]:
  

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.

• [http://partsregistry.org/Part:BBa_K092100 BBa_K092100] :
  

Constitutive RhlR Receiver

This part has been successfully sequenced.

• [http://partsregistry.org/Part:BBa_K092200 BBa_K092200] :

RhlI with RBS and terminator

• [http://partsregistry.org/Part:BBa_K092300 BBa_K092300] :

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.

• [http://partsregistry.org/Part:BBa_K092400 BBa_K092400] :
  

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

• [http://partsregistry.org/Part:BBa_K092600 BBa_K092600] :
  

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.

• [http://partsregistry.org/Part:BBa_K092700 BBa_K092700] :

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) [http://partsregistry.org/Part:BBa_K092000 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.

• [http://partsregistry.org/Part:BBa_K092800 BBa_K092800] :
  

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 LacI_m : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4

• [http://partsregistry.org/Part:BBa_K092900 BBa_K092900] :

rhlR controlled by a constitutive promoter, and lacI_m with a RhlR promoter.

• [http://partsregistry.org/Part:BBa_K092901 BBa_K092901]: Project Part

Sequence containing : - rhlR controlled by a constitutive promoter - lacI_m 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.

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