Plasmid

Part:BBa_K4630202

Designed by: Shuangwu Wu   Group: iGEM23_WHU-China   (2023-10-11)


pCasop-stgRNA(1+2+3)

Though we get a useful pCasop（BBa_K4630201）, we intent to make the entire device more compact and better contribute to the synthetic biology. In detail, we design Golden Gate Assembly primers for ligation of the Cas9 coding sequence and the Lambda Red, araC sequence. Considering the multi-cloning site on the pCDF-Duet1 backbone, we attempt to add XhoI and NotI prefix on the ligated Cas-Lambda sequence. Then the segment can be ligated to the plasmid by restriction cloning. We re-organize them in one integrated plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 679
    Illegal EcoRI site found at 2517
    Illegal EcoRI site found at 6611
    Illegal XbaI site found at 10720
    Illegal SpeI site found at 671
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 679
    Illegal EcoRI site found at 2517
    Illegal EcoRI site found at 6611
    Illegal NheI site found at 2276
    Illegal NheI site found at 9920
    Illegal SpeI site found at 671
    Illegal NotI site found at 696
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 679
    Illegal EcoRI site found at 2517
    Illegal EcoRI site found at 6611
    Illegal BglII site found at 5378
    Illegal BamHI site found at 1043
    Illegal BamHI site found at 4555
    Illegal BamHI site found at 6545
    Illegal BamHI site found at 12241
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 679
    Illegal EcoRI site found at 2517
    Illegal EcoRI site found at 6611
    Illegal XbaI site found at 10720
    Illegal SpeI site found at 671
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 679
    Illegal EcoRI site found at 2517
    Illegal EcoRI site found at 6611
    Illegal XbaI site found at 10720
    Illegal SpeI site found at 671
    Illegal NgoMIV site found at 9173
    Illegal AgeI site found at 878
    Illegal AgeI site found at 6380
    Illegal AgeI site found at 8044
    Illegal AgeI site found at 8920
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 860
    Illegal SapI site found at 6362


[edit]
Categories
Parameters
None