Coding

Part:BBa_K4275008:Design

Designed by: Zheng Xiaoyou   Group: iGEM22_GreatBay_SCIE   (2022-09-29)


FAST-PETase-t


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The sequence of the first 264 amino acids is identical with the free FAST-PETase (BBa_K4275007).

2. A 10 aa long GS linker (GGGGS)2 is appended following the Ser264 residue at the C' terminal of the original sequence.

3. The 79 amino acid long type-I dockerin domain is fused at the terminal of the GS linker, followed by a TEV site and a 8xhis affinity purification tag (HHHHHHHH).

4. DNA sequence is codon-optimized based on the codon-usage table of E.coli Strain K12.MG1655

Source

Ideonella sakaiensis
Clostridium thermocellum (Dockerin-I domain)

References

1. Lu, Hongyuan et al. "Machine Learning-Aided Engineering Of Hydrolases For PET Depolymerization". Nature, vol 604, no. 7907, 2022, pp. 662-667. Springer Science And Business Media LLC, https://doi.org/10.1038/s41586-022-04599-z.