Coding
CrCPR

Part:BBa_K4947029:Design

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


C. roseus Cytochrome P450 Reductase Codon-Optimized CDS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1408
    Illegal PstI site found at 821
    Illegal PstI site found at 1465
    Illegal PstI site found at 1629
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1408
    Illegal PstI site found at 821
    Illegal PstI site found at 1465
    Illegal PstI site found at 1629
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1408
    Illegal BamHI site found at 1299
    Illegal XhoI site found at 6
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1408
    Illegal PstI site found at 821
    Illegal PstI site found at 1465
    Illegal PstI site found at 1629
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1408
    Illegal PstI site found at 821
    Illegal PstI site found at 1465
    Illegal PstI site found at 1629
    Illegal NgoMIV site found at 981
    Illegal NgoMIV site found at 1674
  • 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 CPR was chosen rationally after thorough literature review. CPR is an enzyme involved in the biosynthetic pathway of daidzein, helping convert liquiritigenin into 2,7,4’-trihydroxyisoflavanone with the help of isoflavone synthase. The gene sequence was sourced from NCBI GenBank [1], and produced by Twist Bioscience.

References

1. https://www.ncbi.nlm.nih.gov/protein/Q05001.1

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

5. Chemler, et al., J. (2010). A Versatile Microbial System for Biosynthesis of Novel Polyphenols with Altered Estrogen Receptor Binding Activity. Cell. https://www.cell.com/cell-chemical-biology/pdf/S1074-5521(10)00121-3.pdf