Difference between revisions of "Part:BBa K2206003"

Revision as of 22:51, 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-27b-3p 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-27b-3p 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

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 BsaI.rc site found at 758

@@ Line 5: / Line 5: @@
 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-27b-3p 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-27b-3p present.
+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-27b-3p 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-27b-3p present. This part can therefore be used to quantify the levels of hsa-mir-15b-5p.
-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).