Coding

Part:BBa_K2929004:Design

Designed by: James Hammond   Group: iGEM19_St_Andrews   (2019-10-11)
Revision as of 15:15, 11 October 2019 by Registry (Talk | contribs)

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mOrange + SpyTag


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 820
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 594
    Illegal AgeI site found at 706
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Besides codon-optimising the sequence for our E. coli chassis, we used a protein tertiary structural predictor to assess if the protein folds correctly. The predictor suggested that the protein folds correctly, having a similar topology to mOrange (PDB ID: 2H5O). This suggested to us that the SpyTag would have no negative impact on the fluorescence of the part.


Source

Obtained the sequences for BBa_E2050 and BBa_K1159201 from the registry, then joined the sequences via a 2xGGSG linker and ordered sequence from IDT. The sequence was codon-optimised for E. coli using the IDT codon optimisation tool.

References