Part:BBa_J58100:Design
AND-type promoter synergistically activated by cI and CRP
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 11
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 11
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 11
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We take the lambda Prm promoter sequence from the Registry (BBa_I12007), and we added the CRP binding site upstream of that promoter. Following Joung et al. the centered position of the cI operator was fixed to -42, and for the CRP to -93.5. This forced us to do a lot of cut-and-paste of the original Prm sequence to create (presumably) harmless spacers between the operator regions. Unfortunately the CRP consensus binding site contained a XbaI site. Further work is needed to do mutagenesis on this site while preserving the binding affinity.
Source
The sources of this promoter are, on the one hand, from the bacteriophage lambda, and, on the other hand, from the E. coli. We take the lambda Prm promoter (modified to be activated but not repressed by cI, part ref. BBa_I12007), and the binding site region for CRP from E. coli.
References
JK Joung, DM Koepp, A Hochschild. Synergistic activation of transcription by bacteriophage lambda cI protein and E. coli cAMP receptor protein. Science (1994), 265, 1863-1866.
Bintu et al., Transcriptional regulation by the numbers: applications. Current Opinion in Genetics & Development 2005, 15:125–135