Part:BBa_M36138:Design
Lac Operon modified CAP binding site
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to modify the CAP binding site, we took the original sequence between the LacI and LacZ genes in E coli, and changed the nucleotides the 22 bp known to be the CAP binding site. In order to make sure we sufficiently changed the binding site, we changed all of the purines to pyrimidines and vice versa.. In order to make our construct viable, we added some filler LacI sequence ahead of the intergene sequence. This part is meant to be inserted in the E coli sensor test rig, BBa_M36099, because it contains E coli endogenous genes that are necessary for the function of our part (LacI, CometGFP).
We had to determine where the binding site is, and what endogenous sequences were necessary to include in our construct. For example, we noticed that we should not include a promoter and LacI as it may result in leakage. We also did not include LacZ because we will use CometGFP as part of our sensor. We are trying both a deletion of the binding site and a modification of it just in case the 22 bp length is necessary for replication of the lac operon.
Source
E coli's natural genomic sequence (Ecogene) E coli strain: DH10B