Composite

Part:BBa_K5159029:Design

Designed by: Tianyue WU   Group: iGEM24_HKUST   (2024-10-02)


KU70 Knockout Cassette


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1035
    Illegal PstI site found at 3675
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1035
    Illegal PstI site found at 3675
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1035
    Illegal BglII site found at 3267
    Illegal XhoI site found at 1981
    Illegal XhoI site found at 2010
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1035
    Illegal PstI site found at 3675
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1035
    Illegal PstI site found at 3675
    Illegal NgoMIV site found at 40
    Illegal NgoMIV site found at 1123
    Illegal AgeI site found at 2595
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1641
    Illegal BsaI site found at 1924
    Illegal BsaI.rc site found at 1839
    Illegal SapI site found at 140
    Illegal SapI site found at 2392
    Illegal SapI.rc site found at 718


Design Notes

As before the knockout Yarrowia lipolytica still shows a higher tendency of utilizing NHEJ instead of HR, two 1000 bp homologous arms in length are used, which is necessary for the efficient knockout to occur.


Source

Please refer to the sources of each subparts.

References