Composite
Part:BBa_K3440020
Designed by: Victoria Muliadi, Julie Cordier Group: iGEM20_Stockholm (2020-10-23)
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
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1733
Illegal BamHI site found at 301 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1864
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 776
Illegal BsaI.rc site found at 3249
[edit]
Categories
Parameters
//chassis/prokaryote/ecoli
//classic/signalling/receiver
//function/biosynthesis/ahl
//function/degradation/ahl
//function/reporter
//function/reporter/fluorescence
//classic/signalling/receiver
//function/biosynthesis/ahl
//function/degradation/ahl
//function/reporter
//function/reporter/fluorescence
origin | Aequorea victoria |
output | Oscillating GFP production |
signalling_molecule | QS of Lux system and Rhl system |