Coding
SbpA | Luc

Part:BBa_K525411:Design

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-09-13)

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

  • 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 20(5):895–903.