Coding

Part:BBa_K3602015

Designed by: Flora Fuglsang   Group: iGEM20_SDU-Denmark   (2020-10-23)


Cas12a w. tags, optimized for human cells

This part includes the sequence for the Cas12a protein with the 6x His Tag/Twin Strep/SUMO tags. The sequence is codon-optimized for expression in human cells. The tags can be used for his-tag purification of the protein after expression from e.g. E. coli TOP10.

Cas12a is a protein with endonuclease activity. In complex with a sgRNA, Cas12a cleaves nearby ssDNA upon complementary binding of a target sequence matching the sgRNA. This can be utilized for detection of different targets, as a DNA reporter can be used for the cleavage.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 349
    Illegal PstI site found at 1411
    Illegal PstI site found at 3820
    Illegal PstI site found at 4018
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 349
    Illegal PstI site found at 1411
    Illegal PstI site found at 3820
    Illegal PstI site found at 4018
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 533
    Illegal BglII site found at 2075
    Illegal BglII site found at 2384
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 349
    Illegal PstI site found at 1411
    Illegal PstI site found at 3820
    Illegal PstI site found at 4018
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 349
    Illegal PstI site found at 1411
    Illegal PstI site found at 3820
    Illegal PstI site found at 4018
    Illegal NgoMIV site found at 3619
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 131


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Parameters
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