Coding

Part:BBa_K4275010:Design

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


MHETase-t


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 123
    Illegal NgoMIV site found at 237
    Illegal NgoMIV site found at 693
    Illegal AgeI site found at 190
    Illegal AgeI site found at 307
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The sequence of the first 565 amino acids is identical with the free MHETase (BBa_K4275009).

2. A 10 aa long GS linker (GGGGS)2 is appended following the Pro565 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. Knott, Brandon C. et al. "Characterization And Engineering Of A Two-Enzyme System For Plastics Depolymerization". Proceedings Of The National Academy Of Sciences, vol 117, no. 41, 2020, pp. 25476-25485. Proceedings Of The National Academy Of Sciences, https://doi.org/10.1073/pnas.2006753117.