Regulatory

Part:BBa_K2560302

Designed by: Benjamin Daniel   Group: iGEM18_Marburg   (2018-10-06)


Malonyl-CoA Sensor. This part is a PhytoBrick variant


piGEM2502_fapO

This part contains the fapO sequence, as well as a strong artificial promoter P_23100. FapO is the binding site for the repressor protein FapR. When FapR is present, transcription of the coding sequence controlled by J_23100 is blocked. When FapR binds Malonyl-CoA it leaves the fapO site and transcription can occur.

Usage and Biology

  • This part acts as a promoter controlled by the repressor FapR.
  • The system was taken from B. subtilis 168
  • Malonyl-CoA is a normal metabolite and completely harmless. For induction, a concentration between 2 and 25 mM can be used.
  • We designed this biobrick in order to get a system with a usable readout for the intracellular Malonyl-CoA concentration.
  • Can with be combined with a several reporter genes. We had the best results using the LUX operon.


File:T--Marburg--SensorConstructMalonylCoA-01.png
Figure 1
This is a schematic, illustrating the genetic context and assembly of the Malonyl-CoA sensor



' Figure 1
This is a schematic, illustrating the working mechanism of the Malonyl-CoA sensor


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None