Device

Part:BBa_K1587005:Design

Designed by: Louise GODY   Group: iGEM15_Toulouse   (2015-09-14)

Butyrate synthesis pathway (BBa_1587004) without ccr gene 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 352
    Illegal BamHI site found at 2269
    Illegal XhoI site found at 42
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1535
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 387
    Illegal BsaI.rc site found at 804
    Illegal BsaI.rc site found at 1872
    Illegal SapI.rc site found at 1884

Design Notes

There are two restriction sites of Nhe1 and Xho between p(Bla) and RBS sequence.

Source

We did the assembly of several sequences from ncbi, and hbd and crt genes which came originally from Clostridium acetobutylicum were optimized to work in E.coli.

References

- Louis P & Flint HJ (2009) Diversity, metabolism and microbial ecology of butyrate-producing bacteria from the human large intestine. FEMS Microbiol. Lett. 294: 1–8
- Atsumi S, Cann AF, Connor MR, Shen CR, Smith KM, Brynildsen MP, Chou KJY, Hanai T & Liao JC (2008) Metabolic engineering of Escherichia coli for 1-butanol production. Metabolic Engineering 10: 305–311
- Wallace KK, Bao ZY, Dai H, Digate R, Schuler G, Speedie MK & Reynolds KA (1995) Purification of crotonyl-CoA reductase from Streptomyces collinus and cloning, sequencing and expression of the corresponding gene in Escherichia coli. Eur. J. Biochem. 233: 954–962