Coding

Part:BBa_K4275003:Design

Designed by: Yunyue Wang   Group: iGEM22_GreatBay_SCIE   (2022-09-29)


TaLPMO-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 AgeI site found at 273
    Illegal AgeI site found at 622
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. The catalytic domain and the dockerin domain are interspaced with a glycine-rich linker.

2. 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).

3. DNA sequence is codon-optimized based on the codon-usage table of E.coli Strain K12.MG1655, but the protein was later expressed in K.marxianus due to the lack of post-translational modifications and the formation of inclusion bodies in E.coli BL21(DE3).

Source

Thermoascus aurantiacus
Clostridium thermocellum (Dockerin-I domain)

References

1. Harris, Paul V. et al. "Stimulation Of Lignocellulosic Biomass Hydrolysis By Proteins Of Glycoside Hydrolase Family 61: Structure And Function Of A Large, Enigmatic Family". Biochemistry, vol 49, no. 15, 2010, pp. 3305-3316. American Chemical Society (ACS), https://doi.org/10.1021/bi100009p.