Difference between revisions of "Part:BBa K4035001:Design"
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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The start and stop codon of the CUP1 genomic sequence were removed before insertion in the plasmid in order to fuse the protein to Aga2 and V5.
+
The sequence has been cloned using Gibson Assembly. The start and stop codon of the CUP1 genomic sequence were removed in order to fuse the protein to Aga2 and V5.
  
 
===Source===
 
===Source===

Revision as of 19:09, 21 October 2021


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 sequence has been cloned using Gibson Assembly. 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 metallothionein 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.