Coding

Part:BBa_K1179017:Design

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


A mutant DNA binding Cas9 with an N-terminal linker and BsaI site for ligation


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3823
    Illegal EcoRI site found at 4199
    Illegal PstI site found at 244
    Illegal PstI site found at 2212
    Illegal PstI site found at 2446
    Illegal PstI site found at 3658
    Illegal PstI site found at 3962
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3823
    Illegal EcoRI site found at 4199
    Illegal PstI site found at 244
    Illegal PstI site found at 2212
    Illegal PstI site found at 2446
    Illegal PstI site found at 3658
    Illegal PstI site found at 3962
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3823
    Illegal EcoRI site found at 4199
    Illegal BglII site found at 251
    Illegal BglII site found at 1188
    Illegal BamHI site found at 2006
    Illegal XhoI site found at 3544
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3823
    Illegal EcoRI site found at 4199
    Illegal PstI site found at 244
    Illegal PstI site found at 2212
    Illegal PstI site found at 2446
    Illegal PstI site found at 3658
    Illegal PstI site found at 3962
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3823
    Illegal EcoRI site found at 4199
    Illegal PstI site found at 244
    Illegal PstI site found at 2212
    Illegal PstI site found at 2446
    Illegal PstI site found at 3658
    Illegal PstI site found at 3962
    Illegal NgoMIV site found at 2740
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4175
    Illegal BsaI.rc site found at 4166


Design Notes

Has a BsaI site on the n-terminal part of the Cas9 coding sequence for fusion protein construction.

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 2013 MIT iGEM team.

References