Coding
CFY

Part:BBa_K1381017

Designed by: Alexander Wirtanen, Niklas Handin, Marcus Hong and Jonas Mattisson   Group: iGEM14_Uppsala   (2014-10-03)
Revision as of 13:24, 17 October 2014 by Alexander v (Talk | contribs)

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 and our target pathogene. The Colicin Fy will target the cellmembrane of it's target.

The killing system We designed the killing system to secret the colicin Fy with the help of an export tag, the USP45 BBa_K1381018. This part will be without promoter, RBS and export tag. It only contains the colicin Fy and a 6x His-Tag for characterization. It is designed so that you can easily couple another promoter or export tag of your choice. This part contains the restriction sites NgoMIV att the start of the sequence to more easily fuse proteins like an export tag to it. Since it got a NgoMIV site att the start of the sequence it is possible to fuse other sequence if they follow the Freiburg Biobrick standard 25 and contains an AgeI site. However our part is not entirely built in the Freiburg standard Biobrick 25 since it does not contain an AgeI site.

Characterization To be able to Characterize colicin Fy we had to couple it to a promoter and a RBS, this we did by creating the construct BBa_K1381023. Figure 4: The gel contains the following from the left to the right
Protein ladder
Eluent 1 for lysate produced by bacteriocin producing bacteria
Eluent 3 for lysate produced by bacteriocin producing bacteria
Eluent 2 for lysate produced by bacteriocin producing bacteria
Final eluent test-tube 4 for lysate produced by bacteriocin producing bacteria
Final eluent test-tube 6 for lysate produced by bacteriocin producing bacteria
Eluent 2 for lysate produced by the negative control
Eluent 3 for lysate produced by the negative control
Final eluent test-tube 4 for lysate produced by the negative control
Final eluent test-tube 6 for lysate produced by the negative control

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 4
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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