Coding

Part:BBa_K1962006:Design

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


Truncated Colicin E9 Lacking Bacteriocin Active Domain


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1381
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1375
    Illegal BamHI site found at 1357
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Compliant with RFC[10].


Source

We used the Chimera molecular modelling software to determine the position of the DNase domain of Colicin E9, we then designed a truncated Colicin E9, without this domain. The protein sequence was then back translated into DNA and codon optimised for E. coli K-12 before being synthesised by IDT as a gBlock gene fragment. Using oligonucleotide primers we then amplified the truncated colicin E9 with a multiple cloning site containing the following restriction sites: BamHI, KpnI, SalI, BglII and NheI at the 3' end.

References