Part:BBa_K4947022:Design

G. uralensis 4-Coumarate-CoA Ligase 1 Codon-Optimized CDS

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 766
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 766
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1151
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 766
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 766
  Illegal NgoMIV site found at 1144
  Illegal NgoMIV site found at 1335
  Illegal AgeI site found at 913
  
• 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 4CL was chosen rationally after thorough literature review. 4CL is an enzyme involved in the biosynthetic pathway of daidzein, converting p-coumaric acid into p-coumaroyl-CoA. The gene sequence was sourced from NCBI GenBank [1], and produced by Twist Bioscience.

References

1. https://www.ncbi.nlm.nih.gov/protein/QDG03190.1

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

3. 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/

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