Coding

Part:BBa_K525562:Design

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-10-28)

Fusion protein of NADP+ Oxidoreductase and BisdA and BisdB with middle strong promoter,RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1411
    Illegal BamHI site found at 2149
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 65
    Illegal AgeI site found at 2402
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 746


Design Notes

  • Fusion protein of FNR, BisdA and BisdB behind medium strong constitutive promoter to avoid overexpression which can leed to misfolding as [http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2672.2008.03843.x/full Sasaki et al. (2008)] reported.
  • Fusion protein in and assembled via Freiburg BioBrick assembly standard (RFC 25) leaving AgeI and NgoMIV restriction sites


Source

  • bisdA and bisdB originated in Sphingomonas bisphenolicum AO1
  • FNR originated in Escherichia coli TOP10
  • All subparts (except the FNR) can be found in the kit plates


References

Sasaki M, Tsuchido T, Matsumura Y (2008) Molecular cloning and characterization of cytochrome P450 and ferredoxin genes involved in bisphenol A degradation in Sphingomonas bisphenolicum strain AO1, [http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2672.2008.03843.x/full J Appl Microbiol 105(4):1158-1169].