Project

Part:BBa_K1774000:Design

Designed by: HKU iGEM 2015   Group: iGEM15_Hong_Kong_HKU   (2015-09-18)

CRISPR-Cas9 contaiment device repressed by arabinose and tryptophan


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 7081
    Illegal SpeI site found at 7089
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1247
    Illegal NheI site found at 6532
    Illegal NheI site found at 6555
    Illegal SpeI site found at 7081
    Illegal SpeI site found at 7089
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1187
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 7081
    Illegal SpeI site found at 7089
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 7081
    Illegal SpeI site found at 7089
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI site found at 5785
    Illegal SapI.rc site found at 6761


Design Notes

The whole construct is synthesized using gBlock DNA synthesis and assembled by Hi-Fi assembly. This prototype containment device is designed specifically to be experiment on E. coli BL21 (DE3), thus it may not work in other strain because of the nature of CRISPR-Cas9.

There are a few illegal sites within the promoter region which may not be able to removed by silent mutation.

Source

Cas9 originated from S. pyogenes SF370

References