Coding

Part:BBa_K3515015:Design

Designed by: Kody Klupt, Sono Khan   Group: iGEM20_Queens_Canada   (2020-06-29)
Revision as of 18:01, 29 June 2020 by Registry (Talk | contribs)

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Alpha-Klotho Binding Protein with cysteine modification(s) and FRET to monitor phosphate levels usin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2337
    Illegal XhoI site found at 1876
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 871
    Illegal AgeI site found at 1698
    Illegal AgeI site found at 2567
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Design Considerations: Added a Cysteine for immobilization (L664C) opposite to the active site region. Removed C223A, C316A, C370N, C673L, C96S, and C970S. to ensure only one cysteine would bind to an immobilization linker arm. Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. All disulfides in Alpha-Klotho were kept to ensure proper structural folding. No modifications were made to the FRET acceptor or donor.


Source

The source of this part is its sequence retrieved from the European Nucleotide Archive (M37722.1) along with our own modifications.

References