Project
SgsE|LigA

Part:BBa_K525005:Design

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

Fusion Protein of S-Layer SgsE and LigA


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
    Illegal BglII site found at 3819
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 4417
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1657
    Illegal SapI.rc site found at 4033


Design Notes

Assembled via Freiburg BioBrick assembly standard (RFC 25) from BBa_K525303 and BBa_K525720


Source

  • 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_K525720 NAD+ dependent DNA ligase from Escherichia coli TOP10


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, 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, FEMS Microbiol Lett 267(2):131-144.

Tang Z, Liu P, Ma C, Yang X, Wang K, Tan W, Lv X (2011) Molecular Beacon Based Bioassay for Highly Sensitive and Selective Detection of Nicotinamide Adenine Dinucleotide and the Activity of Alanine Aminotransferase, Anal Chem 83(7):2505-2510.