Part:BBa_K525582:Design

Fusion protein of NADP+ Oxidoreductase and BisdA and BisdB, polycistronic

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 1449
  Illegal BamHI site found at 2187
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 65
  Illegal NgoMIV site found at 850
  Illegal AgeI site found at 812
  Illegal AgeI site found at 2440
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 746
  

Design Notes

• Polycistronic expression of FNR and fusion protein of BisdA and BisdB.

• Sequence of bisdA and bisdB was not entered correctly into the parts.igem. For further information see our notes in BBa_K123000:Experience and BBa_K123001:Experience

• bisdA and bisdB BioBricks were sent to the parts.igem in the Freiburg BioBrick assembly standard (RFC 25) leading to illegal AgeI and NgoMIV restriction sites in this sequence

Source

• Organism: bisdA and bisdB originated in Sphingomonas bisphenolicum AO1

• DNA (probably) synthesized and with optimated codon-usage for E. coli

• FNR originated in Escherichia coli TOP10

• Anderson promoter BBa_J23110, RBS BBa_B0034, BisdA BBa_K123000 and BBa_K123001 from parts.igem and kit plates, respectively.

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].