Part:BBa_K525411:Design

Fusion Protein of S-Layer SbpA 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 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 4846
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 493
  Illegal BsaI.rc site found at 622
  Illegal SapI.rc site found at 4003
  

Design Notes

• BBa_K525403 fused to BBa_K525999
  • BBa_K525403 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_K525403 fused to BBa_K525999
  • 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_K525999 Firefly luciferase from P. pyralis and Promega pGL3 vector, respectively. Converted into Freiburg BioBrick standard RFC 25 from BioBrick BBa_I712019

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].