Part:BBa_K525002:Design

Fusion protein of SbpA, NADP+ Oxidoreductase, BisdA and BisdB with PT7/lac

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 104
  Illegal BglII site found at 221
  Illegal BamHI site found at 4539
  Illegal BamHI site found at 5277
  Illegal XhoI site found at 1996
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 76
  Illegal AgeI site found at 5530
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 493
  Illegal BsaI.rc site found at 622
  Illegal BsaI.rc site found at 3874
  

Design Notes

Assembled via Freiburg BioBrick assembly standard (RFC 25) from BBa_K525403 and BBa_K525560

Source

• BBa_K525403 synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins

• Promoter: fusion promoter between T7 promoter and lac-operator
  • T7 promoter from T7 phage
  • lac-operator from E. coli

• BBa_K525401 S-layer gene sbpA synthesized, originated in Lysinbacillus sphaericus CCM 2177

• BBa_K525499 Ferredoxin-NADP⁺ oxidoreductase from E. coli TOP10

• BBa_K123000 and BBa_K123001 bisdA and bisdB from Sphingomonas bisphenolicum AO1, codon optimized

References

Badelt-Lichtblau H, Kainz B, Völlenkle C, Egelseer EM, Sleytr UB, Pum D, Ilk N (2009) Genetic Engineering of the S-Layer Protein SbpA of Lysinibacillus sphaericus CCM 2177 for the Generation of Functionalized Nanoarrays, Bioconjugate Chem [http://pubs.acs.org/doi/abs/10.1021/bc800445r 20(5):895–903].

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].

Sleytr UB, Huber C, Ilk N, Pum D, Schuster B, Egelseer EM (2007) S-layers as a tool kit for nanobiotechnological applications, [http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2006.00573.x/full FEMS Microbiol Lett 267(2):131-144].