Part:BBa_K3257101
Cre Recombinase with SsrA(WVLAA)-aTc Inducible
According to our modeling, the number of Cre recombinase in one E.coli cell should be a relatively small one, more specifically 5 per cell, so that mutations will accumulate after rounds of R-Evolution process. Based on this instruction, we have spent much effort in lowering the steady-state expression level of Cre. An important approach is to attach 5 different types of degradation tags after the CDS of Cre. Using EGFP as a reporter, we can see that the degradation rate of degradation tag with the amino acid residue sequence of AANDENWVLAA is moderate for Cre recombinase to reach a steady but low level of expression. Transformed into BL21 (DE3) with another plasmid containing lox sites and analyzed by agarose gel, we have seen that Cre is slightly restricted to knockout DNA fragment between two loxP sites.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 436
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 122
- 1000COMPATIBLE WITH RFC[1000]
| None |