Composite

Part:BBa_K2206005

Designed by: Sammy Lovat   Group: iGEM17_CLSB-UK   (2017-10-02)
Revision as of 22:52, 2 October 2017 by SLovat (Talk | contribs)


Toehold switch for hsa-miR-27b-3p with luciferase

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-27b-3p (the trigger RNA). The toehold switch is activated by hsa-mir-27b-3p and regulates production of Luciferase. Luminescence intensity from Luciferase is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present. This part can therefore be used to quantify the levels of hsa-mir-27b-3p.

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).

N.B. This part contains a strong RBS sequence.

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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 936


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Categories
Parameters
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