Device

Part:BBa_K1065309:Design

Designed by: Fabio Digiacomo   Group: iGEM13_UNITN-Trento   (2013-09-18)
Revision as of 20:59, 3 October 2013 by Cridelbianco (Talk | contribs) (→‎Design Notes)


light regulated circuit producing amilGFP+EFE at dark


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3309
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 770
    Illegal NgoMIV site found at 842
    Illegal NgoMIV site found at 932
    Illegal NgoMIV site found at 950
    Illegal NgoMIV site found at 1462
    Illegal NgoMIV site found at 1755
    Illegal NgoMIV site found at 1849
    Illegal AgeI site found at 484
    Illegal AgeI site found at 1630
    Illegal AgeI site found at 4060
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1519
    Illegal BsaI.rc site found at 383


Design Notes

We designed this part by inserting the part BBa_K1065302 uptsream BBa_K1065002. So we built a part consisting of the blue light circuit that regulates amilCP+EFE production. Note that a terminator is missing before PfixK2. We suggest to instert at least one terminator before the promoter in order to have a better defined switch from the ON state to the OFF state.

Source

The part sources are:
BBa_K1065302 (the improved version of the part BBa_K952003);
BBa_K1065002 (EFE+terminators)

References

1) Moglich A, Ayers RA and Moffat K. (2009) Design and Signaling Mechanism of Light-Regulated Histidine Kinases. J. Mol. Bio. 385, 5, 1433-1444.
2) Ohlendorf, R., Vidavski, R.R., Eldar, A., Moffat, K. & Möglich, A.(2012). From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. J. Mol. Biol., 416: 534: 542
3) Nagahama K, Ogawa T, Fujii T, Tazaki M, Tanase S, et al. (1991) Purification and properties of an ethylene-forming enzyme from Pseudomonas syringae pv. phaseolicola PK2. Journal of General Microbiology 137: 2281–2286.
Fukuda H, Ogawa T, Ishihara K, Fujii T, Nagahama K, et al. (1992) Molecular cloning in Escherichia coli, expression, and nucleotide sequence of the gene for the ethylene-forming enzyme of Pseudomonas syringae pv. phaseolicola PK2. Biochem Biophys Res Commun 188: 826–832.
4) Guerrero F, Carbonell. V., Cossu M, Correddu D, Jones PR (2012) Ethylene Synthesis and Regulated Expression of Recombinant Protein in Synechocystis sp. PCC 6803. PLoS ONE 7(11): e50470.