Plasmid

Part:BBa_K4761108

Designed by: Pinhan Pan   Group: iGEM23_HiZJU-China   (2023-09-30)


pDonor

pDonor used in knock-in of ARO10 and UGT85A1 gene. The carrier for transposon structure. Encodes a mini-transposon derived from V. cholerae CAST system. It is used in CRISPR-Transposon Three-Plasmid system.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1018
    Illegal PstI site found at 828
    Illegal PstI site found at 865
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1018
    Illegal PstI site found at 828
    Illegal PstI site found at 865
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1018
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1018
    Illegal PstI site found at 828
    Illegal PstI site found at 865
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1018
    Illegal PstI site found at 828
    Illegal PstI site found at 865
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 850
    Illegal BsaI.rc site found at 412
    Illegal SapI.rc site found at 152


Characterization

Installation of ARO10(BBa_K4761000) and UGT85A1(BBa_K4761002) into pDonor has been proved successful. By using the CRISPR-Transposon system all the two genes have been successfully knocked in the genome of DH5α.

Fig Verification of successfully constructed pDonor-ARO10 and pDonor-UGT85A1 plasmids. Ready for gene integration.
[edit]
Categories
Parameters
None