Coding

Part:BBa_K1179057:Design

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


VN173-Cas9(D10A and H840A mutant)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4401
    Illegal EcoRI site found at 4762
    Illegal PstI site found at 243
    Illegal PstI site found at 822
    Illegal PstI site found at 2790
    Illegal PstI site found at 3024
    Illegal PstI site found at 4236
    Illegal PstI site found at 4540
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4401
    Illegal EcoRI site found at 4762
    Illegal PstI site found at 243
    Illegal PstI site found at 822
    Illegal PstI site found at 2790
    Illegal PstI site found at 3024
    Illegal PstI site found at 4236
    Illegal PstI site found at 4540
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4401
    Illegal EcoRI site found at 4762
    Illegal BglII site found at 829
    Illegal BglII site found at 1766
    Illegal BamHI site found at 1
    Illegal BamHI site found at 2584
    Illegal XhoI site found at 4122
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4401
    Illegal EcoRI site found at 4762
    Illegal PstI site found at 243
    Illegal PstI site found at 822
    Illegal PstI site found at 2790
    Illegal PstI site found at 3024
    Illegal PstI site found at 4236
    Illegal PstI site found at 4540
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4401
    Illegal EcoRI site found at 4762
    Illegal PstI site found at 243
    Illegal PstI site found at 822
    Illegal PstI site found at 2790
    Illegal PstI site found at 3024
    Illegal PstI site found at 4236
    Illegal PstI site found at 4540
    Illegal NgoMIV site found at 3318
    Illegal AgeI site found at 4734
  • 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