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Composite
SgsE | YFP

Part:BBa_K525305:Design

Designed by: Timo Wolf   Group: iGEM11_Bielefeld-Germany   (2011-09-10)

Fusion Protein of S-Layer SgsE and mCitrine


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 3121
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1657


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_K525301 S-layer gene sgsE synthesized, originated in Geobacillus stearothermophilus NRS 2004/3a
  • 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].