Coding
SgsE | Red
Part:BBa_K525312:Design
Designed by: Anna Drong Group: iGEM11_Bielefeld-Germany (2011-09-11)
Fusion Protein of S-Layer SgsE and Oxidoreductase
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 167
Illegal BglII site found at 1022 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 76
Illegal AgeI site found at 3148 - 1000INCOMPATIBLE 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].