Part:BBa_K525312:Design

Fusion Protein of S-Layer SgsE and Oxidoreductase

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 3148
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1657
Illegal BsaI.rc site found at 3082

Design Notes

BBa_K525303 fused to BBa_K525499
- BBa_K525303 synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
- BBa_K525499 from Escherichia coli TOP10 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_K525303 fused to BBa_K525499
- 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_K525499 Ferredoxin-NADP⁺ oxidoreductase from E. 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, [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].