Coding

Part:BBa_K1962002:Design

Designed by: Frank Sargent   Group: iGEM16_Dundee   (2016-10-11)


Truncated Colicin Ia Lacking Bacteriocin Active Domain


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1375
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1369
    Illegal BamHI site found at 1351
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

REC[10] compliant


Source

We used the Chimera molecular modelling software to determine the sequence of the cytotoxic domain of Colicin iA, we then designed a truncated Colicin Ia, without this domain. The sequence was then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment. Using primers we amplified the truncated Colicin Ia with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI located at the 3' end. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminal end of the truncated colicin and thereby generating synthetic colicins.

References