Composite

Part:BBa_K3218003

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

The complex of dCas9 and FokI allows for extra specificity when inducing mutations in a targeted manner. The FokI is fused to the dCas9 and can only cleave DNA when it forms a dime with another FokI. For this reason we created introns (BBa_K3218000) that can be inserted into gene of interest and create a very targeted region of mutations using two dCas9-FokI complexes. The gene is under the control of the divergent inducible promoter GAL1-10 induced by galactose.

Sequence and Features


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]


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Categories
Parameters
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