Difference between revisions of "Part:BBa K525311:Design"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K525311 short</partinfo>
 
<partinfo>BBa_K525311 short</partinfo>
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===Design Notes===
 
===Design Notes===
−
Assembled
 
  
 +
* <partinfo>K525303</partinfo> fused to <partinfo>K525999</partinfo>
 +
** <partinfo>K525303</partinfo> synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
 +
** <partinfo>K525999</partinfo> from ''Photinus pyralis'' / Promega pGL3 vector 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===
 
===Source===
  
−
BBa_K525303; BBa_K525999
+
* <partinfo>K525303</partinfo> fused to <partinfo>K525999</partinfo>
 +
** <partinfo>K525303</partinfo> 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''
 +
 
 +
* <partinfo>K525301</partinfo> S-layer gene ''sgsE'' synthesized, originated in ''Geobacillus stearothermophilus'' NRS 2004/3a
 +
 
 +
* <partinfo>K525999</partinfo> Firefly luciferase from ''P. pyralis'' and Promega pGL3 vector, respectively. Converted into Freiburg BioBrick standard RFC 25 from BioBrick <partinfo>I712019</partinfo>
 +
 
 +
 
  
 
===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].
 +
 +
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].

Latest revision as of 17:10, 21 September 2011

Fusion Protein of S-Layer SgsE and Firefly-Luciferase


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 4054
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1657
    Illegal SapI.rc site found at 3211


Design Notes

  • BBa_K525303 fused to BBa_K525999
    • BBa_K525303 synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
    • BBa_K525999 from Photinus pyralis / Promega pGL3 vector 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

  • 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_K525999 Firefly luciferase from P. pyralis and Promega pGL3 vector, respectively. Converted into Freiburg BioBrick standard RFC 25 from BioBrick BBa_I712019


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].