Coding

Part:BBa_K1179030:Design

Designed by: Lauren St. Hilaire   Group: iGEM13_MIT   (2013-09-15)


Cas9-VN173 (D10A and H840A mutant)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4748
    Illegal PstI site found at 288
    Illegal PstI site found at 2256
    Illegal PstI site found at 2490
    Illegal PstI site found at 3702
    Illegal PstI site found at 4006
    Illegal PstI site found at 4413
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4748
    Illegal PstI site found at 288
    Illegal PstI site found at 2256
    Illegal PstI site found at 2490
    Illegal PstI site found at 3702
    Illegal PstI site found at 4006
    Illegal PstI site found at 4413
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4748
    Illegal BglII site found at 295
    Illegal BglII site found at 1232
    Illegal BamHI site found at 1
    Illegal BamHI site found at 2050
    Illegal XhoI site found at 3588
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4748
    Illegal PstI site found at 288
    Illegal PstI site found at 2256
    Illegal PstI site found at 2490
    Illegal PstI site found at 3702
    Illegal PstI site found at 4006
    Illegal PstI site found at 4413
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4748
    Illegal PstI site found at 288
    Illegal PstI site found at 2256
    Illegal PstI site found at 2490
    Illegal PstI site found at 3702
    Illegal PstI site found at 4006
    Illegal PstI site found at 4413
    Illegal NgoMIV site found at 2784
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Not applicable

Source

The mutant DNA-binding Cas9 originates from the type II prokaryote CRISPR system in Streptococcus pyogenes. The split venus fluorescent protein is a genetic mutant of the GFP from Aequorea victoria. Constructed by performing a BsaI digestion of an IDT synthesized VN173 gBlock and the Cas9 mutant DNA binding entry vector.

References