Difference between revisions of "Part:BBa K4035001:Design"
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(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. | (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. | ||
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| + | (2) https://www.uniprot.org/uniprot/P0CX80 | ||
Latest revision as of 19:18, 21 October 2021
CUP1 fused to Aga2 and tagged with a V5 epitope
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 319
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 376
Illegal PstI site found at 319 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 562
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 319
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 319
- 1000COMPATIBLE 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 yeast copper metallothionein 1 protein (2) and was inserted in the pCTcon2-V5 (1) plasmid that already contains 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.