Part:BBa_I12035:Design
Modified lamdba Prm promoter (repressed by p22 cI without cooperativity) RBS+
- 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 O-R3 region of the lambda cI strand was mutated by 7 nt to eliminate any binding in the region. This was not replaced with the O-R1 region of p22 due to the fact that the O-R1 site had a length of 18 nt, while the space between the -10 and -35 regions was only 16 nt. Thus, spacing would not be preserved, and this could cause many problems. However, since p22 cI must still repress the production of the lambda cI protein, the OR-1 site of p22 was still placed within the construct. The positioning is right after the -10 region (between the -10 region and the RBS). Thus, when p22 cI binds in, protein production should still be inhibited, whereas, if p22 does not bind in, there will just be a little extra code in the mRNA, which theoretically should not cause any problems.
Source
Bushman, F. D. The bacteriophage 434 right operator roles of O-R1, O-R2, and O-R3. J. Mol. Biol. (1993) 230, 28-40.