Part:BBa_K4475006:Design

CTB3 Gene from Cercospora beticola

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1263
  Illegal XbaI site found at 100
  Illegal XbaI site found at 457
  Illegal XbaI site found at 925
  Illegal XbaI site found at 1243
  Illegal XbaI site found at 1381
  Illegal XbaI site found at 1651
  Illegal XbaI site found at 2104
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1263
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1263
  Illegal BglII site found at 388
  Illegal BglII site found at 562
  Illegal BglII site found at 1291
  Illegal BglII site found at 2563
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1263
  Illegal XbaI site found at 100
  Illegal XbaI site found at 457
  Illegal XbaI site found at 925
  Illegal XbaI site found at 1243
  Illegal XbaI site found at 1381
  Illegal XbaI site found at 1651
  Illegal XbaI site found at 2104
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1263
  Illegal XbaI site found at 100
  Illegal XbaI site found at 457
  Illegal XbaI site found at 925
  Illegal XbaI site found at 1243
  Illegal XbaI site found at 1381
  Illegal XbaI site found at 1651
  Illegal XbaI site found at 2104
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Cut site eligibility for RFC 1000 achieved through silent mutations. N-terminal 6xHis tagged and codon optimized for yeast.

Source

Genome of Cercospora Beticola https://www.ncbi.nlm.nih.gov/gene/35424646

References

Newman AG, Townsend CA. Molecular Characterization of the Cercosporin Biosynthetic Pathway in the Fungal Plant Pathogen Cercospora nicotianae. J Am Chem Soc. 2016 Mar 30;138(12):4219-28. doi: 10.1021/jacs.6b00633. Epub 2016 Mar 16. PMID: 26938470; PMCID: PMC5129747.