![](https://parts.igem.org/images/partbypart/icon_coding.png)
Part:BBa_K4275010:Design
MHETase-t
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.