Part:BBa_K3257101
Cre Recombinase with SsrA(WVLAA)-aTc Inducible
For the best composite part, we would like to present you with our Cre recombinase together with degradation tag (https://2019.igem.org/Team:Fudan-TSI/Part_Collection). Cre recombinase (BBa_K3257044 https://parts.igem.org/Part:BBa_K3257044) is responsible for the recombination process between the reverse-transcribed cDNA and the target Gene of Interest (GOI), which is rather one of the most important processes in our system.
Because of the leakage of uninduced Cre expression, 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 (BBa_E0040 https://parts.igem.org/Part:BBa_E0040) as a reporter, we can see that the degradation rate of degradation tag with the amino acid residue sequence of AANDENWVLAA (BBa_K3257073 https://parts.igem.org/Part:BBa_K3257073) is moderate for Cre recombinase to reach a steady but relatively low level of expression.
Transformed into BL21 (DE3) with another plasmid containing lox sites and analyzed by colony PCR, we can see that homologous recombination happens between the same lox sites instead of gene knockout (Figure 1).
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]
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