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Part:BBa_K3228103:Design

Designed by: Maurice Mager   Group: iGEM19_Marburg   (2019-10-18)


pMC_2_002_cpf1+crRNA-GFP_dropout


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1003
    Illegal PstI site found at 3538
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3538
    Illegal NotI site found at 1
    Illegal NotI site found at 5542
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4701
    Illegal XhoI site found at 1053
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1003
    Illegal PstI site found at 3538
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1003
    Illegal PstI site found at 3538
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is the transcriptional unit of the CRISPR associated nuclease Cas12a from Francisella tularensis ssp. novicida together with a spaceholder for the gRNA region of the crRNA. The dropout is a GFP cassette that can be cut out with the BsmbI restriction enzyme. For genome editing you simply put in your desired gRNA (oligo annealing) via a golden gate reaction.


Source

Golden Gate assembly of parts from the Marburg Collection

References