Difference between revisions of "Part:BBa K1381019"

Latest revision as of 22:44, 17 October 2014

B0034-pelB-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.

PelB
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 export tag for E.coli, pelB, 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. 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 also contains an 6x His-tag for characterisation.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 54
Illegal NgoMIV site found at 76
1000
COMPATIBLE WITH RFC[1000]

@@ Line 6: / Line 6: @@
 '''PelB'''<br> PelB is en export tag that works in ''E.coli''.
-'''The killing system'''<br> We designed the killing system to secret the colicin Fy with the help of an export tag, pelB. The part contains the colicin Fy [https://parts.igem.org/Part:BBa_K1381017 BBa_K1381017], the export tag for ''E.coli'', pelB, and an 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 pelB 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.
+'''The killing system'''<br> We designed the killing system to secret the colicin Fy with the help of an export tag, pelB. The part contains the colicin Fy [https://parts.igem.org/Part:BBa_K1381017 BBa_K1381017], the export tag for ''E.coli'', pelB, 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. 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 also contains an 6x His-tag for characterisation.
 <!-- Add more about the biology of this part here