Composite

Part:BBa_K2557031:Design

Designed by: Ge JT   Group: iGEM18_NAU-CHINA   (2018-10-09)


miniCMV promoter-Anti-GFP-mnotch-TEV protease-NLS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 455
    Illegal PstI site found at 499
    Illegal PstI site found at 818
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 62
    Illegal PstI site found at 455
    Illegal PstI site found at 499
    Illegal PstI site found at 818
    Illegal NotI site found at 1616
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2109
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 455
    Illegal PstI site found at 499
    Illegal PstI site found at 818
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 455
    Illegal PstI site found at 499
    Illegal PstI site found at 818
    Illegal NgoMIV site found at 254
    Illegal NgoMIV site found at 959
    Illegal NgoMIV site found at 1136
    Illegal NgoMIV site found at 1674
    Illegal AgeI site found at 71
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1295


Design Notes

Since we need TEV protease to enter the nucleus, we added SV40 NLS to the back of the TEV protease.Since we used animal cells as the chassis organism, the coding sequence was humanized.


Source

We borrowed the Anti-GFP-mnotch-tetR-VP64-V2 plasmid from Fudan University. After the TEV protease sequence used by the iGEM17_Oxford group was found on the iGEM website, it was sent to Kingsray for synthesis.

References