Coding
GuCHR1

Part:BBa_K4947025:Design

Designed by: Williams Liu   Group: iGEM23_Yale   (2023-10-12)


G. uralensis Chalcone Reductase 1 Codon-Optimized CDS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 826
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 826
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 746
    Illegal BglII site found at 830
    Illegal BglII site found at 878
    Illegal XhoI site found at 337
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 826
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 826
    Illegal NgoMIV site found at 555
  • 1000
    COMPATIBLE 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. https://www.ncbi.nlm.nih.gov/nuccore/MK341786.1/

2. View our contributions page (https://2023.igem.wiki/yale/contribution) for a spreadsheet of all our sources!

3. Yan, Y. (2020). De novo biosynthesis of liquiritin in Saccharomyces cerevisiae. NCBI. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7161706/

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

5. Li, J. (2021, September 25). Diversion of metabolic flux towards 5-deoxy(iso)flavonoid production via enzyme self-assembly in escherichia coli. Metabolic Engineering Communications. https://www.sciencedirect.com/science/article/pii/S2214030121000250#bib64