Composite

Part:BBa_K3218003:Design

Designed by: Abraham Grosman   Group: iGEM19_MSP-Maastricht   (2019-10-15)


dCas9-FokI, an endonuclease fused to a deactivated Cas9 under the control of GAL1-10


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4939
    Illegal NheI site found at 5494
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2484
    Illegal BglII site found at 2943
    Illegal BglII site found at 3032
    Illegal BglII site found at 3677
    Illegal BamHI site found at 669
    Illegal XhoI site found at 696
    Illegal XhoI site found at 5472
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1206
    Illegal AgeI site found at 286
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

For this mutation mechanism when designing the introns to be inserted, we had to be sure the at the PAM sequences were out and the FokI endonucelase would be facing the interior of the two introns.


Source

Both dCas9 and FokI come form bacterial genomes.

References