Composite

Part:BBa_K2751010:Design

Designed by: Hao-Tung Liu   Group: iGEM18_NYMU-Taipei   (2018-10-09)


Ubc9-YPet


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 40
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 529
    Illegal XhoI site found at 1258
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1181


Design Notes

This part applies the cutting sites in the prefix of part BBa_K2751008 to insert the Ubc9. The Ubc9 with proper cuttings sites at both ends are generated by PCR. Afterwards, the sequence is inserted into the prefix of part BBa_K2751008 through ligation. As a result, this part contains a ribosome binding site, Ubc9, YPet, and his tag, allowing it to be produced once it is under control of a promoter and purified with Ni2+ chromatography.


Source

Ubc9 DNA sample(pcDNA-T7-Ubc9) is obtained from Dr. Pei-Ching Chang of NYMU and YPet is from Dr. Yaw-Kuen Li of NCTU-Taiwan.

References

1. Song, Y., & Liao, J. (2012, August). An in vitro Förster resonance energy transfer-based high-throughput screening assay for inhibitors of protein-protein interactions in SUMOylation pathway. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/22192309