Part:BBa_K3484004
AHL sfGFP+ASV reporter
This composite part produces sfGFP when a large enough concentration of lactone (AHL) is present in the medium. In this case, it constitutively produces LuxR, which binds to the lactone present in the medium and together, they form a complex that activates the pLux promoter, generating the sfGFP whose fluorescence can be measured. The schematics of this composite part can be seen in Figure 1.
This system allows a fast report on the AHL concentrations in the medium, something important to know the amount of protein. The composite part is Biobrick compatible.
This composite part includes all the following parts:
(BBa_B0015): A reliable double terminator that consists on BBa_B0010 and BBa_B0012.
(BBa_B0033): A weak RBS. The RBS.4 (derivative of BBa_0030).
(BBa_B0032): A medium RBS. The RBS.3 (medium) (derivative of BBa_0030)
(BBa_R0062): LuxR & HSL regulated promoter.
(BBa_J72005): Ptet Promoter. Is activated by the AHL-LuxR complex
(BBa_K3484006): sfGFP+ASV coding sequence
(BBa_B0015) followed by the sfGFP with the ASV degradation tag (This reporter sequence BBa_K3484006) . A control with the original sequence (BBa_I746916), with the same promoter, double terminator and RBS, was done to see how the tag affected the final fluorescence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1249
Illegal AgeI site found at 1372 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1142
None |