Difference between revisions of "Part:BBa K2206003"
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<partinfo>BBa_K2206003 short</partinfo> | <partinfo>BBa_K2206003 short</partinfo> | ||
− | 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, | + | 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. |
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+ | 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 of GFP is proportional to the number of toehold switches activated, thus indicating the levels of hsa-mir-15b-5p present. | ||
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+ | This part can therefore be used to quantify the levels of hsa-mir-15b-5p. | ||
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+ | 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). | ||
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+ | N.B. This part contains a strong RBS sequence. | ||
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Revision as of 22:45, 2 October 2017
Toehold switch for hsa-miR-27b-3p with GFPmut3b
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 GFPmut3b. Fluorescence intensity of GFP 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-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).
N.B. This part contains a strong RBS sequence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 758