Difference between revisions of "Part:BBa K2206002"
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This part codes for a toehold switch that contains a region that is complementary to the microRNA hsa-miR-15b-5p (the trigger RNA). The toehold switch is activated by hsa-miR-15b-5p and regulates production of GFPmut3b. The fluorescence intensity from GFPmut3b is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of GFPmut3b is produced), thus indicating the levels of hsa-miR-15b-5p present (as the more microRNA there is, the greater the number of switches activated). This part can therefore be used to quantify the levels of hsa-miR-15b-5p. | This part codes for a toehold switch that contains a region that is complementary to the microRNA hsa-miR-15b-5p (the trigger RNA). The toehold switch is activated by hsa-miR-15b-5p and regulates production of GFPmut3b. The fluorescence intensity from GFPmut3b is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of GFPmut3b is produced), thus indicating the levels of hsa-miR-15b-5p present (as the more microRNA there is, the greater the number of switches activated). This part can therefore be used to quantify the levels of hsa-miR-15b-5p. | ||
| − | The transcripts levels produced by this part are toxic to E. coli. Therefore, we recommend the use of an inducible non-leaky promoter to allow for amplification in E. coli. For reference, we found BBa_K808000 suitable for | + | The transcripts levels produced by this part are toxic to E. coli. Therefore, we recommend the use of an inducible non-leaky promoter to allow for amplification in E. coli. For reference, we found BBa_K808000 was a suitable replacement for BBa_J23111. |
This part contains a strong RBS sequence. | This part contains a strong RBS sequence. | ||
Latest revision as of 22:51, 30 October 2017
Toehold switch for hsa-miR-15b-5p 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 microRNA hsa-miR-15b-5p (the trigger RNA). The toehold switch is activated by hsa-miR-15b-5p and regulates production of GFPmut3b. The fluorescence intensity from GFPmut3b is proportional to the number of toehold switches activated (as the more switches activated, the greater the amount of GFPmut3b is produced), thus indicating the levels of hsa-miR-15b-5p present (as the more microRNA there is, the greater the number of switches activated). This part can therefore be used to quantify the levels of hsa-miR-15b-5p.
The transcripts levels produced by this part are toxic to E. coli. Therefore, we recommend the use of an inducible non-leaky promoter to allow for amplification in E. coli. For reference, we found BBa_K808000 was a suitable replacement for BBa_J23111.
This part contains a strong RBS sequence.
NUPACK Structure Analysis
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 764