Difference between revisions of "Part:BBa K1381020"

Latest revision as of 22:43, 17 October 2014

J23106-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 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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 95
Illegal NgoMIV site found at 117
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 is coupled with an Anderson promoter, the J23106. It is a fairly strong Anderson promoter. 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 contains a 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 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.
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