Part:BBa_K1913025:Design
Wild type plac-FixK2 hybrid promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to make Fixk2 promoter as an inducible promoter for the toggle switch,we had to add additional repressor operator sequences into the Fixk2 sequence, they are both on upstream and downstream the Fixk2 sequence, which could be bonded by tetrameric Lac repressor resulting in the formation of a DNA loop and consequently on transcription repression.
Source
Genomic sequence of Bradyrhizobium japonicum
References
Oehler S, Eismann E R, Krämer H, et al. The three operators of the lac operon cooperate in repression[J]. The EMBO journal, 1990, 9(4): 973. Nellen-Anthamatten D, Rossi P, Preisig O, et al. Bradyrhizobium japonicumFixK2, a Crucial Distributor in the FixLJ-Dependent Regulatory Cascade for Control of Genes Inducible by Low Oxygen Levels[J]. Journal of bacteriology, 1998, 180(19): 5251-5255.
Mesa S, Ucurum Z, Hennecke H, et al. Transcription activation in vitro by the Bradyrhizobium japonicum regulatory protein FixK2[J]. Journal of bacteriology, 2005, 187(10): 3329-3338.
Ferrières L, Kahn D. Two distinct classes of FixJ binding sites defined by in vitro selection[J]. FEBS letters, 2002, 517(1-3): 185-189.