Part:BBa_K1381022
B0034-USP45-CFY-X6 his
Colicin Fy
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.
USP45
USP45 is en export tag that works in Lactobacillus as well as E.coli.
The killing system
We designed the killing system to secret the colicin Fy with the help of an export tag, USP45. The part contains the colicin Fy BBa_K1381017, the secretion tag USP45, and a RBS. This part do not have any promoter. It is desgined that way so it can be easier to couple a promoter of your choice with the rest of the killing system. However due to problems with the restriction enzymes for AgeI and MgoMIV the Uppsala iGEM team 2014 could not couple the export tag, pelB and the colicin Fy together with the Freiburg Biobrick standard 25. Instead we used the regular Biobrick standard 10 to couple them, which results in a stop codon between USP45 and CFY. For this part to work, the stop codon must be removed with mutagenesis. This part also contains an 6x His-tag for characterisation.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
Illegal NgoMIV site found at 76 - 1000COMPATIBLE WITH RFC[1000]
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