Part:BBa_K3440020

Oscillation circuit under GFP reporter

Pconst(BBa_J23100)-RBS(BBa_B0034)-RhlR(BBa_C0071)-DT(BBa_B0015)-Prhl(BBa_R0071)-RBS(BBa_B0034)-RhlI(BBa_C0070)-RBS(BBa_B0034)-AiiA(BBa_C0060)-RBS(BBa_B0034)-GFP(BBa_E0040)-DT(BBa_B0015)

Usage and Biology

This part produces RhlR constitutively, which can induce Prhl in the presence of C4-HSL lactone. With C4-HSL (or due to the leakiness of Prhl), RhlI, AiiA and GFP are produced under Prhl. RhlI produces C4-HSL, which creates a positive feedback loop on the promoter. However, AiiA degrades C4-HSL, which creates a negative feedback loop on Prhl. Thus, GFP expression oscillates and this part can be used as a reporter to check that the expected oscillations appear.

Theoretical expectations

[[File: T--Stockholm--Modeling.png|thumb|center|1000px|Improved inducible oscillatory circuit at different induction points of 3OC6-HSL. Concentrations of RhlI (I), AiiA (A), MtrB (X), C4-HSL (H), 3OC6-HSL (H2) and LuxR (R) plotted over time] 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
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1733
  Illegal BamHI site found at 301
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1864
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 776
  Illegal BsaI.rc site found at 3249