Difference between revisions of "Part:BBa K3440020"

Revision as of 07:47, 25 October 2020

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

@@ Line 11: / Line 11: @@
 ==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]
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 <span class='h3bb'>Sequence and Features</span>