Composite
Part:BBa_K4275041:Design
Designed by: Wei Zijian Group: iGEM22_GreatBay_SCIE (2022-10-12)
pT7-RBS-CipA1B2C-tT7
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2073
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 354
Illegal XhoI site found at 1277 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 583
Illegal AgeI site found at 365
Illegal AgeI site found at 709
Illegal AgeI site found at 908
Illegal AgeI site found at 1579 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
1. A 6x His affinity purification tag(HHHHHH) is added to N-terminal following the start codon, to allow for Ni-NTA purification.
2. The number of cohesins is cut from 9 to 2, and the number of CBM is cut from 2 to 1 to reduce the metabolic burden and increase the probability of successful expression in the E.coli BL21 recombinant protein expression system.
Source
Clostridium thermocellum
References
1. Anandharaj, Marimuthu et al. "Constructing A Yeast To Express The Largest Cellulosome Complex On The Cell Surface". Proceedings Of The National Academy Of Sciences, vol 117, no. 5, 2020, pp. 2385-2394. Proceedings Of The National Academy Of Sciences, https://doi.org/10.1073/pnas.1916529117.