Team:EPF-Lausanne/Parts

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Cloning Scheme

First plasmid : the basic network components

Second plasmid : the testing component

Parts submitted to the registry

[1] BBa_K092000 :
Composition : B0034 + C0040 + B0010 + B0012 = RBS + TetR with LVA + stop + stop
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].

[2] BBa_K092100 :

Composition : I14033 + B0034 + C0171 + B0010 + B0012 = Constutive promoter + RBS + RhlR + stop + stop
Constitutive RhlR Receiver

[3] BBa_K092200 :

Composition : B0034 + C0070 + B0010 + B0012 = RBS + RhlI + stop + stop
RhlI with RBS and terminator

[4] BBa_K092300 :

Composition : R0040 + B0034 + E1010 = TetR repressible promoter + RBS + RFP
RFP controlled by a TetR promoter : Coding sequence of the RFP with a ribosome binding site and pTetR as promoter. Thus, the transcription of RFP is inhibited when TetR is present.

[5] BBa_K092400 : New part created by 2-steps PCR. RBS (B0034) + LuxI + Terminator (B0010+B0012) :

Coding sequence of the Lux Inducer with a ribosome binding site and terminator. This biobrick is an alternative to the F1610 which didn't work

[6] BBa_K092600 :

Composition : B0034 + C0040 + B0010 + B0012 + R0040 + B0034 + E1010 = RBS + TetR with LVA + stop + stop + TetR repressible promoter + RBS + RFP

TetR cds with RBS and Terminator + pTetR, 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.

[7] BBa_K092700 :

Composition : B0034 + C0040 + B0010 + B0012 + R0040 + B0034 + E1010 + K092400 = RBS + TetR with LVA + stop + stop + TetR repressible promoter + RBS + RFP + RBS +LuxI + stop+stop

TetR cds with RBS and Terminator + pTetR, 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.

[8] BBa_K092800:

Coding sequence for LacI modified with a different rate of translation than LacI, RBS.

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

[9] BBa_K092900:

Composition : I14033 + I1466 + R0071 + K092800 = Constutive promoter + RBS + RhlR + Terminator + RBS + LacIm

RhlR controlled by a constitutive promoter, and LacI modified with a Rhl R promoter.

[10] BBa_K092901: Project Part

Composition : I14033 + I1466 + R0071 + K092800 + B0015 + R0010 + B0034 + C0040 + B0010 + B0012 + R0040 + B0034 + E1010 + K092400

Sequence containing : - RhlR controlled by a constitutive promoter - LacI 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.