Difference between revisions of "Part:BBa K4035002:Design"

 
Line 7: Line 7:
  
 
===Design Notes===
 
===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.  
+
The start and stop codon of the CUP1 genomic sequences were removed in order to fuse the proteins together with the linker.  
  
  
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===Source===
 
===Source===
  
−
The CUP1 sequence is the genomic sequence of the copper metallotionein 1 protein. After having linked two copies with a linker in between, the full sequence was codon optimized by the software of the company that synthetized the sequence in order to avoid loops during syntethis. The CUP1-linker-CUP1 sequence was then inserted in the pCTcon2_V5 plasmid that was already containing Aga2, V5 tag and the Gal1 promoter by the Gibson assembly method.  
+
The CUP1 sequence is the genomic sequence of the copper metallotionein 1 protein. After having linked two copies with a linker in between, the full sequence was codon optimized by the software of the company that synthetized the sequence 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.  
  
 
===References===
 
===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.

Revision as of 05:09, 13 September 2021


Dimerization of the copper metallothionein 1 : CUP1-(GGGGS)3-CUP1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 190
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 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.


Source

The CUP1 sequence is the genomic sequence of the copper metallotionein 1 protein. After having linked two copies with a linker in between, the full sequence was codon optimized by the software of the company that synthetized the sequence 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.

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.