Regulatory
Part:BBa_K3634018:Design
Designed by: Laurence Seeley Group: iGEM20_St_Andrews (2020-08-08)
PL8-UV5
Assembly Compatibility:
- 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
The wt sequence for the lac operon regulatory region was taken from Kalnins A. (1993) [GenBank accession no. J01636]. Within the CAP binding site, bases -66 (G) and -55 (C) of the wt binding region were substituted with A and T respectively to prevent binding of the CAP protein at low glucose concentrations. The wt -10 promoter sequence was also mutated from TATGTT to TATAAT in order to allow σ factor (RpoD) to bind without relying on further activation by the CAP protein (Reznikoff et al., 1978).