Coding

Part:BBa_K4035001:Design

Designed by: Anissa Hammi   Group: iGEM21_EPFL   (2021-08-30)
Revision as of 09:31, 20 October 2021 by Hammi (Talk | contribs) (→‎Source)


CUP1 fused to Aga2 and tagged with a V5 epitope


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 319
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 376
    Illegal PstI site found at 319
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 562
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 319
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 319
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The start and stop codon of the CUP1 genomic sequence were removed in order to fuse the protein to Aga2 and V5.


Source

The CUP1 (BBa_M45090) sequence is the genomic sequence of the copper metallotionein 1 protein and was inserted in the pCTcon2_V5 plasmid that was already containing Aga2, V5 tag and the Gal1 promoter.

References

(1) Chao, G.; Lau, W. L.; Hackel, B. J.; Sazinsky, S. L.; Lippow, S. M.; Wittrup, K. D. Isolating and Engineering Human Antibodies Using Yeast Surface Display. Nat Protoc 2006, 1 (2), 755–768. https://doi.org/10.1038/nprot.2006.94.