Part:BBa_K525512:Design

Polycistronic expression of BisdA and BisdB

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 896
  Illegal BamHI site found at 1284
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 65
  Illegal NgoMIV site found at 421
  Illegal AgeI site found at 383
  Illegal AgeI site found at 1693
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 256
  Illegal BsaI site found at 292
  Illegal BsaI site found at 1503
  

Design Notes

• Polycistronic expression 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

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

• All subparts can be found on one of 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].