Team:ESBS-Strasbourg/27 August 2008

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DryLab

Modeling

WetLab

The verification PCR has shown that the assemble of:

(LexA_Linker2)_(VP16_Linker2)
(LexA_Linker2)_(TUP1_Linker2)
(LacI_Linker2)_(VP16_Linker2)
(LacI_Linker2)_(TUP1_Linker2)
(CI_Linker2)_(VP16_Linker2)
(CI_Linker2)_(TUP1_Linker2)
(tetR_Linker2)_(VP16_Linker2)
(tetR_Linker2)_(TUP1_Linker2)
(mOrange_Linker2)_(mOrange_Linker2)
(mCherry_Linker2)_(mCherry_Linker2)
(CFP_Linker2)_(CFP_Linker2)
(EYFP_Linker2)_(EYFP_Linker2)
LexAo4

was successful.

We ordered primers to amplify the Adh1_terminator out of the genome from S. cerevisiae.

General

We notice that the BioBrick J63001, that we used as template for our EYFP_BB is marked as inconsient.
So we will start again with the BBa_E2030.

We decided to test which order of BD, AD and XFP is best on the example of LexA.

Katja: I found out that 200-300 bp between the operator sites and the TATA box might be fine. So we will use our Cin8 BB (228 bp)as spacer between the operator sites and the cyc promoters. In the same time I saw that in the plasmid on which we base the hypothesis that the strenght of LexAo8 corresponds to CIo3, there is no space between the single konsensus sequences. While we are working with a space of 16 bp between the operator sites. So we will have to see if this works out as well.