Difference between revisions of "Part:BBa K525405:Design"

(→‎Source)
(→‎References)
Line 31: Line 31:
  
 
===References===
 
===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].
+
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].
−
 
+
−
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].
+

Revision as of 11:32, 15 September 2011

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

  • 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_J18931 yellow fluorescent protein (YFP) mCitrine
    • from parts.igem
    • was synthesized before it was sent in

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].