Part:BBa_K5091008:Design

pmsC

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 112
  Illegal PstI site found at 493
  Illegal PstI site found at 871
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 112
  Illegal PstI site found at 493
  Illegal PstI site found at 871
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 654
  Illegal BamHI site found at 852
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 112
  Illegal PstI site found at 493
  Illegal PstI site found at 871
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 112
  Illegal PstI site found at 493
  Illegal PstI site found at 871
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Since pmsC functions alongside pmsB in the biosynthesis of salicylic acid, co-expression of both genes is essential. A single operon with a shared promoter and ribosome binding sites (RBS) for each gene could simplify expression, but balancing the expression levels of both enzymes may require adjusting the RBS strengths.

Additionally, choosing an appropriate promoter, such as an inducible one, can help control the timing and levels of gene expression to prevent metabolic stress on the host organism. Codon optimization may also be necessary if expressing pmsC in a host other than Pseudomonas fluorescens, ensuring efficient translation and protein production. Lastly, to avoid negative effects from salicylic acid overproduction, users should implement a system to monitor salicylic acid levels during the project.

Source

Pseudomonas fluorescens A506, sequence found through Genebank https://www.ncbi.nlm.nih.gov/gene/12961196

References

Pelludat, C., Brem, D., & Heesemann, J. (2003). Irp9, encoded by the high-pathogenicity island of Yersinia enterocolitica, is able to convert chorismate into salicylate, the precursor of the siderophore yersiniabactin. Journal of bacteriology, 185(18), 5648–5653. https://doi.org/10.1128/JB.185.18.5648-5653.2003