Part:BBa_K2206008
Toehold switch for hsa-miR-15b-5p with luciferase and inducible promoter
Toehold switches are synthetic riboregulators that regulate gene expression post-transcriptionally. Gene expression can be activated in the presence of a cognate single stranded RNA molecule that contains an arbitrary sequence (the trigger RNA). The trigger RNA binds to the switch through base pairing, causing a conformational change that results in translation of the downstream protein coding region.
This part codes for a toehold switch that contains a region that is complementary to the micro-RNA hsa-mir-15b-5p (the trigger RNA). The toehold switch is activated by hsa-mir-15b-5p and regulates production of Luciferase. The luminescence intensity of Luciferase is proportional to the number of toehold switches activated (as the greater the number of switches activated, the more luciferase is produced), thus indicating the levels of hsa-mir-15b-5p present. This part can therefore be used to quantify the levels of hsa-mir-15b-5p.
We believe that the transcripts produced from this part are toxic when expressed at reasonably high concentrations in E.coli (e.g. the concentrations produced from the BBa_J23111 constitutive promoter). We therefore redesigned the constructs to contain an inducible promoter to allow for amplification in E.coli.
N.B. This part contains a strong RBS sequence
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Illegal SapI.rc site found at 2102
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