Reporter

Part:BBa_K2525008

Designed by: Kshitij Rai   Group: iGEM17_IIT_Delhi   (2017-10-27)


PsrpR + RBS + GFP + T

This part contains the SrpR promoter regulating the expression of GFP, which was used as a reporter. SrpR 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 3.2. This part was characterized by Voigt et al by conversion into a not gate, by placing the SrpR 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 SrpR 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, SrpR can give a much better and steeper curve than most traditional repressors. This is highly desirable.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 20
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 738


[edit]
Categories
Parameters
None