Part:BBa_K649205
lox71
To characterize this part, we constructed the generator part BBa_K649202.
The lox sequences, lox71 and lox66, have 5 bp on the 5 and 3 ends changed, respectively. DNA segment flanked by lox71 and lox66 marks the point which the enzyme Cre will excise. This Part is expected to express GFP when the lox sites are excised and RFP when they are not. The Cre-mediated recombination of this BioBrick had been studied and proved to be working.
The leftmost is a negative control which don't have Cre-expressing plasmid.
The center is an arabinose induced sample which has both Cre plasmid and BioBrick BBa_K649201.
The rightmost is a uninduced strain which has both plasmid like as the center.
In in vivo assay, arabinose induced strain which has Cre-expressing plasmid(PBAD/araC-Cre, BBa_I718008) was expressing GFP, while negative control which doesn't have Cre plasmid was expressing Red florescence. It means that DNA recombination did happen by Cre recombinase. The uninduced sample was supposed to unexpress Cre recombinase, but because of the leaking of PBAD\araC promoter, the excision occured in almost every cell in medium.
For more information, see [http://2011.igem.org/Team:Tokyo_Tech/Projects/RPS-game/assay#8. our work in Tokyo_Tech 2011 wiki].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| n/a | lox71 |