Device
A112

Part:BBa_K4947036:Design

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


Daidzein Biosynthetic Pathway Module A112


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 908
    Illegal EcoRI site found at 1064
    Illegal XbaI site found at 298
    Illegal XbaI site found at 2465
    Illegal XbaI site found at 3933
    Illegal XbaI site found at 5189
    Illegal SpeI site found at 1
    Illegal SpeI site found at 706
    Illegal PstI site found at 1137
    Illegal PstI site found at 1220
    Illegal PstI site found at 5048
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 908
    Illegal EcoRI site found at 1064
    Illegal NheI site found at 2562
    Illegal SpeI site found at 1
    Illegal SpeI site found at 706
    Illegal PstI site found at 1137
    Illegal PstI site found at 1220
    Illegal PstI site found at 5048
    Illegal NotI site found at 233
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 908
    Illegal EcoRI site found at 1064
    Illegal BglII site found at 4968
    Illegal BglII site found at 5052
    Illegal BglII site found at 5100
    Illegal XhoI site found at 4559
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 908
    Illegal EcoRI site found at 1064
    Illegal XbaI site found at 298
    Illegal XbaI site found at 2465
    Illegal XbaI site found at 3933
    Illegal XbaI site found at 5189
    Illegal SpeI site found at 1
    Illegal SpeI site found at 706
    Illegal PstI site found at 1137
    Illegal PstI site found at 1220
    Illegal PstI site found at 5048
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 908
    Illegal EcoRI site found at 1064
    Illegal XbaI site found at 298
    Illegal XbaI site found at 2465
    Illegal XbaI site found at 3933
    Illegal XbaI site found at 5189
    Illegal SpeI site found at 1
    Illegal SpeI site found at 706
    Illegal PstI site found at 1137
    Illegal PstI site found at 1220
    Illegal PstI site found at 5048
    Illegal NgoMIV site found at 1835
    Illegal NgoMIV site found at 2167
    Illegal NgoMIV site found at 2246
    Illegal NgoMIV site found at 4777
    Illegal AgeI site found at 322
    Illegal AgeI site found at 944
    Illegal AgeI site found at 1400
  • 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

These gene homologs were chosen rationally after thorough literature review. 4CL converts p-coumaric acid into p-coumaroyl-CoA. CHS and CHR convert p-coumaroyl-CoA into isoliquiritigenin. The gene sequences were sourced from NCBI GenBank (view references on the individual parts’ pages!), and produced by Twist Bioscience.

References

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

2. 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

3. Li, J., et al. (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

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. Yan, Y., et al. (2020). De novo biosynthesis of liquiritin in Saccharomyces cerevisiae. Acta Pharmaceutica Sinica B. NCBI. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7161706/