The CFY is a bacteriocin in the colicin family. It is produced naturally by Y.frederiksenii and target close relatives to that bacteria. One among them is the Y.enterocolitica, which is our target pathogen. The colicin Fy will target the cell membrane of its target.
PelB is en export tag that works in E.coli.
The killing system
We designed the killing system to secret the colicin Fy with the help of an export tag, pelB. The part contains the colicin Fy BBa_K1381017, the secretion tag for E.coli pelB and a RBS. This part is coupled with an Anderson promoter, the J23106. It is a fairly strong Anderson promoter. We used the regular Biobrick standard 10 to couple them which results in a stop codon between pelB and CFY. For this part to work, the stop codon must be removed with mutagenesis. This part contains a 6x His-tag for characterisation.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12INCOMPATIBLE WITH RFCIllegal NheI site found at 7
Illegal NheI site found at 30
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal NgoMIV site found at 95
Illegal NgoMIV site found at 117
- 1000COMPATIBLE WITH RFC