Part:BBa_K4947024:Design
G. max Chalcone Reductase 5 Codon-Optimized CDS
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 427
Illegal PstI site found at 310 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 427
Illegal PstI site found at 310 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 427
Illegal XhoI site found at 337 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 427
Illegal PstI site found at 310 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 427
Illegal PstI site found at 310
Illegal NgoMIV site found at 303
Illegal NgoMIV site found at 555 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The codon optimization and domestication was done to improve recombinant expression in E. coli and enable restriction enzyme-based swapping of promoters and terminators, respectively.
Source
This gene homolog that encodes for CHR was chosen rationally after thorough literature review. CHR is an enzyme involved in the biosynthetic pathway of daidzein, converting p-coumaroyl-CoA into isoliquiritigenin with the help of Chalcone Synthase. It is the enzyme that determines whether the pathway proceeds toward genistein or daidzein as a product. The gene sequence was sourced from NCBI GenBank [1], and produced by Twist Bioscience.
References
1. NCBI Reference Sequence: NP_001304585.2
2. View our contributions page (https://2023.igem.wiki/yale/contribution) for a spreadsheet of all our sources!
3. Liu, Q., Liu, Y., Li, G., Savolainen, O., Chen, Y., & Nielsen, J. (2021, October 19). De novo biosynthesis of bioactive isoflavonoids by engineered yeast cell factories. Nature News. https://www.nature.com/articles/s41467-021-26361-1
4. Cross-kingdom expression of synthetic genetic elements promotes discovery of metabolites in the human microbiome. Patel JR, Oh J, Wang S, Crawford JM, Isaacs FJ. Cell. 2022 Apr 28;185(9):1487-1505.e14. doi: 10.1016/j.cell.2022.03.008. Epub 2022 Apr 1. 10.1016/j.cell.2022.03.008 PubMed 35366417