Part:BBa_K1587004:Design

Butyrate synthesis pathway regulated by constitutive promoter p(Bla)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 36
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1717
  Illegal BamHI site found at 3634
  Illegal XhoI site found at 1407
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 332
  Illegal AgeI site found at 1254
  Illegal AgeI site found at 2900
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1752
  Illegal BsaI.rc site found at 2169
  Illegal BsaI.rc site found at 3237
  Illegal SapI site found at 123
  Illegal SapI.rc site found at 3249
  

Design Notes

p(Bla) is followed by NheI restriction site and ccr is followed by XhoI restriction site

Source

Sequences are from Clostridium acetobutylicum :
- hbd http://www.ncbi.nlm.nih.gov/nuccore/NC_003030.1?report=fasta&from=2831733&to=2832581&strand=true
- crt http://www.ncbi.nlm.nih.gov/nuccore/NC_003030.1?report=fasta&from=2835666&to=2836451&strand=true

Sequences are from E. coli
- atoB http://www.ncbi.nlm.nih.gov/nuccore/NC_020990.1?report=fasta&from=6738960&to=6740186&strand=true
- tesB http://www.ncbi.nlm.nih.gov/nuccore/NC_000913.3?report=fasta&from=474301&to=475161&strand=true

Sequence is from Streptomyces caulinus :
- ccr

The strong RBS and Terminator were used according to respectively BBa_B0030 and BBa_B1006

References

- El-Hussiny A, J. Bacteriol, 2013. Effect of an Oxygen-Tolerant Bifurcating Butyryl Coenzyme A Dehydrogenase/Electron-Transferring Flavoprotein Complex from Clostridium difficile on Butyrate Production in Escherichia coli.
- Mukesh S, J Agric Food Chem, 2014. Metabolic Engineering of Escherichia coli for Production of Butyric Acid
- Atsumi S, 2008, Metab Eng. Metabolic engineering of Escherichia coli for 1-butanol production. Metabolic Engineering 10:305–311.