Part:BBa_K343007:Design
Photosensor generator
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1863
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 153
Illegal NgoMIV site found at 411
Illegal AgeI site found at 1665 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1107
Illegal BsaI.rc site found at 1380
Illegal SapI site found at 881
Illegal SapI.rc site found at 1881
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
Sensory Rhodopsin II and HtrII domains were taken from the genome of Natronomonas Pharaonis. The Tar methyl accepting chemotaxis protein was taken from Salmonella Enterica Serovar Typhimurium. The synthetic linker and fusing of the HAMP domains of HtrII and StTar was done by the team behind the referenced article (1).
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.