Generator
pUS203

Part:BBa_K1388000:Design

Designed by: Rokiah Alford, Tom Geddes, Andreas Bachler   Group: iGEM14_USyd-Australia   (2014-10-07)


IntI1 coding region under AraC-pBad control


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1312
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1223
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1058
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1557
    Illegal SapI site found at 1040


Design Notes

The IntI1 code we found from genomic sequences had prohibited iGEM restriction sites so the sequence was modified to make it compliant with iGEM standards. We ordered the modified sequence as a gBlock (a ~1.1 Kbp DNA block) and was inserted into the AraC-pBad/plasmid backbone with Gibson Assembly. The sequence we made was further modified to include DNA to make Gibson Assembly easier. As Integrase can be toxic to the cell in high concentrations ligation of the BioBrick into a low-copy plasmid is highly recommended and the iGEM prefix and suffix should facilitate this process.

Source

This part is built up from the BBa_K731201 iGEM plasmid (which has AraC-pBad) and the code for the IntI1 enzyme came from a genomic source.

References