Part:BBa_K4947008

RBS for Gu4CL1, expressed in E. coli

This is the ribosome binding sequence for G. uralensis 4-Coumarate-CoA ligase 1.

Description

This is the ribosome binding sequence for G. uralensis 4-Coumarate-CoA ligase 1. This was designed using the CAD-SGE algorithm designed by Jaymin Patel in Farren Isaacs’ lab at Yale University. The algorithm was run repeatedly until the translation initiation rate reached over fifty thousand.

Usage

This part was used in the assembly of the biosynthetic pathway of daidzein. In the construct, it preceded the CDS for G. uralensis 4-Coumarate-CoA ligase 1. This part, along with the rest of the biosynthetic pathway of daidzein, was successfully constructed, cloned into, and integrated into the genome of E. coli DH10Beta.

Experience

This part had little to no difficulty in amplification. It also was an effective ribosome binding site.

References

1. 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 Algorithm: https://cad-sge.com/

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]