Coding

Part:BBa_K4170005:Design

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


Lbucas13a assembly part 4


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal suffix found in sequence at 617
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 618
    Illegal PstI site found at 632
    Illegal NotI site found at 625
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 262
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal suffix found in sequence at 618
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 618
    Illegal PstI site found at 632
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2
    Illegal BsaI.rc site found at 640


Design Notes

This part derives from pGJK_His-SUMO-LbuCas13a plasmid (Plasmid #172488) by mutagenesis PCR to efficiently mutate illegal restriction sites which hinder compatibility with RFC [10] iGEM standard. In addition, the primers exploited incorporate BsaI recognition sites external of the amplified sequence to allow for efficient Golden Gate assembly with the parts 1-3 and the linearilized plasmid backbone.


Source

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

References