Part:BBa_K525311:Design

Fusion Protein of S-Layer SgsE and Firefly-Luciferase

Assembly Compatibility:

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

  Illegal BglII site found at 167
  Illegal BglII site found at 1022
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 76
  Illegal AgeI site found at 4054
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1657
  Illegal SapI.rc site found at 3211
  

Design Notes

• BBa_K525303 fused to BBa_K525999
  • BBa_K525303 synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
  • BBa_K525999 from Photinus pyralis / Promega pGL3 vector 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_K525303 fused to BBa_K525999
  • BBa_K525303 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_K525301 S-layer gene sgsE synthesized, originated in Geobacillus stearothermophilus NRS 2004/3a

• BBa_K525999 Firefly luciferase from P. pyralis and Promega pGL3 vector, respectively. Converted into Freiburg BioBrick standard RFC 25 from BioBrick BBa_I712019

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].