Plasmid

Part:BBa_K4630200

Designed by: Shuangwu Wu   Group: iGEM23_WHU-China   (2023-10-09)


plasmid pCas for expressing Cas9 and Lambda Red recombinases

Based on a previous study, we adopted a one-plasmid system which we called pCas (pRed-Cas9-poxb300) that combines Cas9 and Lambda-Red recombinases in E. coli, which reported to achieve 100% genome editing efficiency in 6 hours of induction. This new composite part consists of a Cas9 coding sequence (BBa_K1218011), and a Lambda-Red recombinases coding sequence (BBa_K1433005). The expression of Cas9 and Lambda-Red is under the control of araBAD promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1344
    Illegal SpeI site found at 5563
    Illegal PstI site found at 11241
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1344
    Illegal NheI site found at 1103
    Illegal SpeI site found at 5563
    Illegal PstI site found at 11241
    Illegal NotI site found at 5882
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1344
    Illegal BglII site found at 4205
    Illegal BamHI site found at 3382
    Illegal BamHI site found at 7647
    Illegal BamHI site found at 11830
    Illegal XhoI site found at 4323
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1344
    Illegal SpeI site found at 5563
    Illegal PstI site found at 11241
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1344
    Illegal SpeI site found at 5563
    Illegal PstI site found at 11241
    Illegal NgoMIV site found at 10790
    Illegal AgeI site found at 7482
    Illegal AgeI site found at 9139
    Illegal AgeI site found at 11665
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 7464
    Illegal SapI site found at 10730
    Illegal SapI site found at 10940
    Illegal SapI site found at 11647


[edit]
Categories
Parameters
None