Composite

Part:BBa_K2019002:Design

Designed by: John Lazar   Group: iGEM16_Northwestern   (2016-10-12)


mRFP + sgRNA targeting mRFP Coding Region (saCas9)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 1099
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1076
    Illegal SpeI site found at 1099
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 1099
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 1099
    Illegal AgeI site found at 781
    Illegal AgeI site found at 893
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Considering that this construct would have to be cotransformed into a cell line with saCas9 expressed in the BBa_R0040-PSB1C3 backbone, all tests of this construct were in the PSB1T3 backbone.


Source

The guide insert was designed using Benchling's gRNA design tool.

References

F. Ann Ran, Le Cong, Winston X. Yan, David A. Scott, Jonathan S. Gootenberg, Andrea J. Kriz, Bernd Zetsche, Ophir Shalem, Xuebing Wu, Kira S. Makarova, Eugene V. Koonin, Phillip A. Sharp, Feng Zhang. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015;520.