Difference between revisions of "Part:BBa K2206006"

 
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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, resulting in translation of the 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 GFPmut3b. Fluorescence intensity from GFP is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present.
 
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, resulting in translation of the 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 GFPmut3b. Fluorescence intensity from GFP is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present.
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We believe that the transcripts produced by 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 us to amplify our constructs in E.coli.  
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We believe that the transcripts produced by 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.  
  
  

Revision as of 20:31, 2 October 2017


Toehold switch for hsa-miR-15b-5p with GFPmut3b 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, resulting in translation of the 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 GFPmut3b. Fluorescence intensity from GFP is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present. We believe that the transcripts produced by 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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1938
    Illegal SapI site found at 961