Composite

Part:BBa_K3738027:Design

Designed by: Emily Hagens and Mark Lea   Group: iGEM21_Lethbridge   (2021-10-21)
Revision as of 23:07, 21 October 2021 by Registry (Talk | contribs)

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Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6xHistidine Tag and crRNA


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 443
    Illegal BglII site found at 1379
    Illegal BglII site found at 1589
    Illegal BglII site found at 1643
    Illegal BglII site found at 1874
    Illegal BglII site found at 2147
    Illegal BglII site found at 2633
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
    Illegal NgoMIV site found at 2095
    Illegal NgoMIV site found at 2731
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part has been codon optimized for E.coli. The part must be in vitro transcribed to obtain the direct-repeat stem loop RNA structure and complex with Cas13a.


Source

The Mcy gene cluster comes from Microcystic Aeruginosa, however the crRNA originates from Leptrotrichia buccalis. The Cas13a protein comes from the Gram-negative bacteria Leptotrichia buccalis (Lbu).

References