Part:BBa_K343003:Design

NpSopII-NpHtrII-StTar (M-fusion)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 1783
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 73
  Illegal NgoMIV site found at 331
  Illegal AgeI site found at 1585
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1027
  Illegal BsaI.rc site found at 1300
  Illegal SapI site found at 801
  Illegal SapI.rc site found at 1801
  

Design Notes

NpSopII-NpHtrII-StTar fusion-chimera protein sequence from Jung K-H, Spudich EN, Trivedi VD and Spudich JL (1).

The first 224 amino acid residues come from the NpSopII gene, encoding a bluelight photon receptor with 15 residues removed at the C-terminal end. The following 9 amino acids are a linker. the last part is HtrII fused with Tsr from E.Coli at the HAMP domain. The complex' first 125 amino acid residues come from HtrII and the remaining 279 from Tsr.

The constitutive promoter R0040 was chosen for its medium strength, so that there would be a rather large amount of the fusion, chimera protein expressed, but not so much that it would be lethal to the cell. The easy repression and inhibition of repression through TetR and tetracycline makes for an easily controlled expression if that is needed. The doible terminator B0015 was chosen for it's reliability and good reviews on the parts.igem.

Source

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References

[http://jb.asm.org/cgi/content/full/183/21/6365] Jung K-H, Spudich EN, Trivedi VD and Spudich JL ; An Archaeal Photosignal-Transducing Module Mediates Phototaxis in Escherichia coli; Journal of Bacteriology, Nov. 2001, p. 6365–6371.