DNA

Part:BBa_K2429011:Design

Designed by: Nia Myrie   Group: iGEM17_MIT   (2017-10-13)
Revision as of 04:46, 27 October 2017 by Nmyrie (Talk | contribs) (→‎Source)


pENTR L. shahii NL-dCas13a-NL


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4422
    Illegal PstI site found at 1903
    Illegal PstI site found at 2272
    Illegal PstI site found at 3001
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4422
    Illegal PstI site found at 1903
    Illegal PstI site found at 2272
    Illegal PstI site found at 3001
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4422
    Illegal BglII site found at 647
    Illegal BglII site found at 1271
    Illegal BglII site found at 1667
    Illegal BglII site found at 1958
    Illegal BglII site found at 2048
    Illegal BglII site found at 3209
    Illegal BglII site found at 3296
    Illegal BamHI site found at 1
    Illegal BamHI site found at 70
    Illegal BamHI site found at 4291
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4422
    Illegal PstI site found at 1903
    Illegal PstI site found at 2272
    Illegal PstI site found at 3001
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4422
    Illegal PstI site found at 1903
    Illegal PstI site found at 2272
    Illegal PstI site found at 3001
    Illegal NgoMIV site found at 4248
    Illegal NgoMIV site found at 4267
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 633
    Illegal SapI.rc site found at 739
    Illegal SapI.rc site found at 1609
    Illegal SapI.rc site found at 2443


Design Notes

This basic part entry vector is flanked by L1 and L2 sites, which are used to denote a gene. This basic part entry vector is flanked by L1 and L2 sites, which are used to denote a gene. There are glycine-serine link between the dCas13a protein and the nuclear localization sequences.

Source

The catalytically active sequence of the protein came from L. shahii Cas13a on Addgene(https://www.addgene.org/79150/), which is also where our team received the original plasmid from. See reference for nuclear localization amino acid sequence.

References