Part:BBa_K525405:Design

Fusion Protein of S-Layer SbpA and mCitrine

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 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 3913
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

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

Design Notes

• BBa_K525403 fused to BBa_J18931
  • BBa_K525403 synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins

• NgoMIV restriction site downstream the start codon to easily fuse an N-terminal domain to the coding sequence

• AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence

Source

• BBa_K525403 fused to BBa_J18931
  • 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

• BBa_J18931 yellow fluorescent protein (YFP) mCitrine
  • from parts.igem
  • was synthesized before it was sent in

References

Kainz B, Steiner K, Möller M, Pum D, Schäffer C, Sleytr UB, Toca-Herrera JL (2010) Absorption, Steady-State Fluorescence, Fluorescence Lifetime, and 2D Self-Assembly Properties of Engineered Fluorescent S-Layer Fusion Proteins of Geobacillus stearothermophilus NRS 2004/3a, [http://pubs.acs.org/doi/abs/10.1021/bm901071b Biomacromolecules 11(1):207-214].

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].