Coding

Part:BBa_K1179054:Design

Designed by: Brandon Nadres   Group: iGEM13_MIT   (2013-09-15)


VN155-Cas9(D10A and H840A mutant)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4347
    Illegal EcoRI site found at 4708
    Illegal PstI site found at 243
    Illegal PstI site found at 768
    Illegal PstI site found at 2736
    Illegal PstI site found at 2970
    Illegal PstI site found at 4182
    Illegal PstI site found at 4486
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4347
    Illegal EcoRI site found at 4708
    Illegal PstI site found at 243
    Illegal PstI site found at 768
    Illegal PstI site found at 2736
    Illegal PstI site found at 2970
    Illegal PstI site found at 4182
    Illegal PstI site found at 4486
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4347
    Illegal EcoRI site found at 4708
    Illegal BglII site found at 775
    Illegal BglII site found at 1712
    Illegal BamHI site found at 1
    Illegal BamHI site found at 2530
    Illegal XhoI site found at 4068
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4347
    Illegal EcoRI site found at 4708
    Illegal PstI site found at 243
    Illegal PstI site found at 768
    Illegal PstI site found at 2736
    Illegal PstI site found at 2970
    Illegal PstI site found at 4182
    Illegal PstI site found at 4486
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4347
    Illegal EcoRI site found at 4708
    Illegal PstI site found at 243
    Illegal PstI site found at 768
    Illegal PstI site found at 2736
    Illegal PstI site found at 2970
    Illegal PstI site found at 4182
    Illegal PstI site found at 4486
    Illegal NgoMIV site found at 3264
    Illegal AgeI site found at 4680
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Not applicable

Source

Was constructed by digestion and ligation of BsaI sites on IDT synthesized split venus proteins and the DNA binding mutant Cas9 entry vector with N-terminal BsaI site. The Cas9 comes from the type II prokaryote CRISPR system in Streptococcus pyogenes. The split venus fluorescence protein is a genetic mutant of GFP which comes from the jellyfish Aequorea victoria.

References