Coding

Part:BBa_K2429025:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-24)
Revision as of 23:31, 24 October 2017 by Nmyrie (Talk | contribs) (→‎Design Notes)

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phEF1a L. shahii Cas13a-DDX6


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 7017
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3049
    Illegal PstI site found at 3418
    Illegal PstI site found at 4147
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 7017
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3049
    Illegal PstI site found at 3418
    Illegal PstI site found at 4147
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 7017
    Illegal BglII site found at 569
    Illegal BglII site found at 1793
    Illegal BglII site found at 2417
    Illegal BglII site found at 2813
    Illegal BglII site found at 3104
    Illegal BglII site found at 3194
    Illegal BglII site found at 4355
    Illegal BamHI site found at 1183
    Illegal BamHI site found at 5437
    Illegal BamHI site found at 5539
    Illegal XhoI site found at 968
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 7017
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3049
    Illegal PstI site found at 3418
    Illegal PstI site found at 4147
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 7017
    Illegal PstI site found at 315
    Illegal PstI site found at 820
    Illegal PstI site found at 3049
    Illegal PstI site found at 3418
    Illegal PstI site found at 4147
    Illegal NgoMIV site found at 703
    Illegal NgoMIV site found at 5394
    Illegal NgoMIV site found at 5413
    Illegal AgeI site found at 81
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1779
    Illegal SapI.rc site found at 1885
    Illegal SapI.rc site found at 2755
    Illegal SapI.rc site found at 3589
    Illegal SapI.rc site found at 6895


Design Notes

This part is Human codon optimized. Furthermore, this specific variation of the protein is catalytically active in bacteria; however, since we used it in mammalian cells, the protein's endonuclease activity isn't expected to be seen. Additionally, the focus is on the protein's binding and blocking ability rather than the catalytic activity.

Source

Sequence for L. shahii Cas13a was found from Addgene(https://www.addgene.org/79150/), which is also where our team received the original plasmid from. DDX6 domain came from another plasmid

References