Coding

Part:BBa_K5094004:Design

Designed by: HuiChing Kuo   Group: iGEM24_KCIS-Xiugang-Taipei   (2024-09-20)


The IsPETase Thr116Ala/Met154Thr, double mutations on the amino acids 116 and 154 in the IsPETase ge


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 220
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The IsPETase gene has several DNA repeat regions so it might be difficult to create a specific amino acid mutation via site-directed mutagenesis


Source

The team created the IsPETase Thr116Ala/Met154Thr, double mutations on the amino acids 116 and 154 in the IsPETase gene, amino acid 116 from the polar uncharged side chain of the Threonine switched to the hydrophobic side chain of the alanine, and amino acid 154 from the hydrophobic side chain of the methionine to the polar uncharged side chain of the Threonine via site-directed mutagenesis

References