- Testing bricks in lab to make sure they work
- Switching parts
- Transfer curve
- Constitutive promoter for LacI: which strains have it?
-R0053 from p22 cII -R0040 from TetR -Z0251 from T7 R0040 is stronger than R0053.
- Summary: contains the sequence for LacI, the CAP binding site, lacI repressor binding site. There are variation in the sequence between different strains. A hard copy of the sequence is in the iGEM binder (it's red and kept by Annie).
A Genetic Switch: Phage Lambda Revisited by Mark Ptashne
- cI and cro are adjacent on the lambda chromosome and transcribed in opposite directions (divergently).
- cI promoter = Prm points polymerase leftward.
- Both promoters do not overlap.
- The production of lacI
- -Components: constitutive promoter, RBS + gene sequence + terminator of LacI gene
- -Test constructs:
- assemble R0053 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
- assemble R0040 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
- assemble Z0251 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
- assemble R0053 + P0412 (LacI gene)
- assemble R0040 + P0412 (LacI gene)
- assemble Z0251 + P0412 (LacI gene)
Qualitative - Quantitative -
- The production of cI/TetR
- -Components: CAP binding site, promoter regulated by LacI, RBS + gene sequence + terminator of cI/TetR gene
- -Test contructs:
- R0010 + E0040 (GFP) or E0433 (LacZ delta m15 bacterial strain)
- R0010 + P0451 (cI without a cI regulated promoter)
- R0010 + Q04510 + E0040/E0433 (Q04510 is cI with a cI regulated promoter)
- R0010 + P0440 (TetR without a TetR regulated promoter)
- R0010 + Q04400 + E0040/E0433 (Q04400 is TetR with a TetR regulated promoter)