Plasmid

Part:BBa_K3602021:Design

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


Cas12a composite part, optimized for E. coli K12


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2312
    Illegal BamHI site found at 1557
    Illegal XhoI site found at 1192
    Illegal XhoI site found at 1594
    Illegal XhoI site found at 3055
    Illegal XhoI site found at 3373
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 886
    Illegal AgeI site found at 2377
    Illegal AgeI site found at 3775
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 949


Design Notes

A positively regulated promoter (BBa_R0011) was used since Cas13a can be toxic to a cell. With the promoter protein production can be regulated so the expressing bacteria will not die.


Source

This sequence is from the PhyLife group of the University of Southern Denmark, where it was originally used for human cells.

References