Difference between revisions of "Part:BBa K1381021"

 
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<partinfo>BBa_K1381021 short</partinfo>
 
<partinfo>BBa_K1381021 short</partinfo>
  
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Promotor - RBS - Export - Bacteriocin - His tag
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'''Colicin Fy'''<br> 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.
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'''PelB'''<br> PelB is en export tag that works in ''E.coli''.
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'''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 J23116. 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. 
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1381021 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1381020 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1381021 parameters</partinfo>
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<partinfo>BBa_K1381020 parameters</partinfo>
 
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Revision as of 21:54, 17 October 2014

J23116-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 an RBS. This part is coupled with an Anderson promoter, the J23116. 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.

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]