Difference between revisions of "Part:BBa K2996704"
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To facilitate measurements, we used a reporter plasmid, with the mRFP gene under the control of a weak promoter_J23117 that is preceded by a sequence rich in NGG PAM sequences on the NT strand. We designed sgRNA that complements the NT strand from at least 20nt upstream from -55 point (With the transcription start denoted +1, RNAP complex covers from -55 to +20.) | To facilitate measurements, we used a reporter plasmid, with the mRFP gene under the control of a weak promoter_J23117 that is preceded by a sequence rich in NGG PAM sequences on the NT strand. We designed sgRNA that complements the NT strand from at least 20nt upstream from -55 point (With the transcription start denoted +1, RNAP complex covers from -55 to +20.) | ||
| − | When the dcas9-sgRNA complex binds to target sequence, RpoA will recruit RNA polymerase, promotor J23117 will be activated. Then, by measuring the biological activity of the downstream reporter gene, we will know the effectiveness of this transcription activation device under different conditions. | + | <br>When the dcas9-sgRNA complex binds to target sequence, RpoA will recruit RNA polymerase, promotor J23117 will be activated. Then, by measuring the biological activity of the downstream reporter gene, we will know the effectiveness of this transcription activation device under different conditions. |
Check BBa_K2996706 for more details. | Check BBa_K2996706 for more details. | ||
Latest revision as of 23:39, 21 October 2019
target sequence upstream of promotor
To facilitate measurements, we used a reporter plasmid, with the mRFP gene under the control of a weak promoter_J23117 that is preceded by a sequence rich in NGG PAM sequences on the NT strand. We designed sgRNA that complements the NT strand from at least 20nt upstream from -55 point (With the transcription start denoted +1, RNAP complex covers from -55 to +20.)
When the dcas9-sgRNA complex binds to target sequence, RpoA will recruit RNA polymerase, promotor J23117 will be activated. Then, by measuring the biological activity of the downstream reporter gene, we will know the effectiveness of this transcription activation device under different conditions.
Check BBa_K2996706 for more details.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 52
Illegal NheI site found at 75 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 39
- 1000COMPATIBLE WITH RFC[1000]