Composite

Part:BBa_K3782027:Design

Designed by: Jana Näf   Group: iGEM21_UNILausanne   (2021-10-20)


Cas9 with sgRNA 3 and donor sequence


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1072
    Illegal BglII site found at 1762
    Illegal BglII site found at 4596
    Illegal BamHI site found at 3404
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4573
    Illegal NgoMIV site found at 5713
    Illegal AgeI site found at 4860
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1679
    Illegal SapI.rc site found at 4406


Design Notes

The sgRNA sequence was designed by finding sequences that are specific to the InaZ gene with low off-target effects. The donor sequence was designed by looking at the genome of Pseudomonas syringae pv. syringae B301D.



Source

pJH1 plasmid The sgRNA and donor sequence were synthesized and added to the plasmid through Golden Gate Assembly and Gibson Assembly.

References