Coding

Part:BBa_K1179008:Design

Designed by: Nelson Hall   Group: iGEM13_MIT   (2013-09-15)


A mutant DNA binding Cas9 with a C-terminal linker and BsaI site for ligation


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4243
    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
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4243
    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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4243
    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 4243
    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
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3867
    Illegal EcoRI site found at 4243
    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 NgoMIV site found at 2784
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4219
    Illegal BsaI.rc site found at 4210


Design Notes

It has a BsaI site at the end of the Cas9 coding sequence for scarless ligation.

Source

The mutant DNA-binding Cas9 originates from the type II prokaryote CRISPR system in Streptococcus pyogenes. Sincere thanks to Samira Kiani for contributing this entry vector to the MIT 2013 iGEM team.

References