Coding

Part:BBa_K4170002:Design

Designed by: Alexandros Giannopoulos Dimitriou   Group: iGEM22_Thessaloniki_Meta   (2022-09-27)


bdSUMO- Lbucas13a assembly part 1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 9
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 9
    Illegal NotI site found at 15
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 9
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 9
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 9
    Illegal XbaI site found at 24
    Illegal AgeI site found at 377
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2
    Illegal BsaI.rc site found at 1234


Design Notes

This part derives from pGJK_His-SUMO-LbuCas13a plasmid (Plasmid #172488) by mutagenesis PCR to efficiently mutate illegal restriction sites. In addition, the primers exploited incorporate BsaI recognition sites external of the amplified sequence to allow for efficient Golden Gate assembly with the parts 2-4 and the linearilized plasmid backbone.


Source

This part derives from pGJK_His-SUMO-LbuCas13a plasmid (Plasmid #172488).

References