Part:BBa_K4947032:Design
T. pratense Isoflavone Synthase Codon-Optimized CDS
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 838
Illegal EcoRI site found at 1237
Illegal PstI site found at 277 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 838
Illegal EcoRI site found at 1237
Illegal PstI site found at 277 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 838
Illegal EcoRI site found at 1237
Illegal BglII site found at 665 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 838
Illegal EcoRI site found at 1237
Illegal PstI site found at 277 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 838
Illegal EcoRI site found at 1237
Illegal PstI site found at 277 - 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 IFS/2-HIS was chosen rationally after thorough literature review. IFS/2-HIS is an enzyme involved in the biosynthetic pathway of daidzein, converting liquiritigenin into 2,7,4’-trihydroxyisoflavanone with the help of cytochrome p450 reductase. The gene sequence was sourced from NCBI GenBank [1], and produced by Twist Bioscience.
References
1. https://www.ncbi.nlm.nih.gov/nuccore/AY253284.1
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