Difference between revisions of "Part:BBa K4035009:Design"

 
 
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===Source===
 
===Source===
  
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The CUP1 sequence is the genomic sequence of the copper metallotionein 1 protein. The linker sequence is the reverse transcription of the GGGGS amino acid sequence. After having designed the dimer sequence and before synthetizing it, the full sequence was codon optimized by the software of the company that synthetized it in order to avoid loops during syntethis. The CUP1-linker-CUP1 sequence was then inserted by Gibson Assembly in the pCTcon2_V5 plasmid containing Aga2, V5 tag and the Gal1 promoter.  
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The CUP1 sequence is the genomic sequence of the yeast copper metallothionein 1 protein (2). The linker sequence is the reverse transcription of the GGGGS amino acid sequence. After having designed the dimer sequence and before synthetizing it, the full sequence was codon optimized by the software of the company that synthetized it to avoid loops during syntethis. The CUP1-linker-CUP1 sequence was then inserted by Gibson Assembly in the pCTcon2-V5 plasmid (1) containing Aga2, V5 tag and the Gal1 promoter.  
  
 
===References===
 
===References===
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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.
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(2) https://www.uniprot.org/uniprot/P0CX80

Latest revision as of 20:50, 21 October 2021


Expression of the CUP1-(GGGGS)3-CUP1 dimer on the extracellular membrane of S. cerevisiae


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1000
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The start and stop codon of the CUP1 genomic sequences were removed in order to fuse the proteins together with the linker, Aga2 and V5.


Source

The CUP1 sequence is the genomic sequence of the yeast copper metallothionein 1 protein (2). The linker sequence is the reverse transcription of the GGGGS amino acid sequence. After having designed the dimer sequence and before synthetizing it, the full sequence was codon optimized by the software of the company that synthetized it to avoid loops during syntethis. The CUP1-linker-CUP1 sequence was then inserted by Gibson Assembly in the pCTcon2-V5 plasmid (1) 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.

(2) https://www.uniprot.org/uniprot/P0CX80