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:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 455
Illegal PstI site found at 499
Illegal PstI site found at 818 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 2109
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 455
Illegal PstI site found at 499
Illegal PstI site found at 818 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.