Coding

Part:BBa_K4035003:Design

Designed by: Anissa Hammi   Group: iGEM21_EPFL   (2021-09-09)
Revision as of 15:50, 9 September 2021 by Registry (Talk | contribs)

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Dimerization of the copper metallothionein 1 : CUP1-(GGGGS)4-CUP1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 90
    Illegal PstI site found at 408
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 90
    Illegal PstI site found at 408
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 235
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 90
    Illegal PstI site found at 408
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 90
    Illegal PstI site found at 408
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 84


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. In addition, the full sequence was codon optimized before being ordered in order to avoid loops formation during synthesis.


Source

The CUP1 sequence is the genomic sequence of the copper metallotionein 1 protein. The sequence of th linker comes from a reverse translation of the amino acid sequence GGGGS and codon optimized for yeast S. cerevisiae. 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.

References