Part:BBa_K2525006
PphlF + RBS + GFP + T
This part contains the PhlF promoter regulating the expression of GFP, which was used as a reporter.
PhlF is a repressor with higher cooperativity than most traditional repressors such as LacI, TetR and cI. This repressor was first reported by Voigt et al (Nature Chem Biol, 2014), where they reported a cooperativity of 4.5. This part was characterized by Voigt et al by conversion into a not gate, by placing the PhlF gene under the pTac promoter. This promoter is inducible by IPTG, and produced increasing levels of PhlF upon increments of IPTG. This was then connected to a node containing the YFP gene under an Orf2 repressible promoter, creating a NOT Gate with IPTG as the input. The input - output curve was plotted to determine the cooperativity.
The high cooperativity of this part is of great significance to synthetic biology, as it can be used to give steeper response curves for repression. Ideally, when we tend to look at genetic elements as digital logic tools, we require steep transitions of protein levels from 0 to 1 or 1 to 0 (representing and ON and OFF state). However, due to various limitations in biological systems, the response is often in the form of a gradient and not a steep curve. Due to its high cooperativity, PhlF can give a much better and steeper curve than most traditional repressors. This is highly desirable.
<h> Chracterization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 719
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