Difference between revisions of "Part:BBa K2996704"
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When the dcas9-sgRNA complex binds to target sequence, RpoA will recruits 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. | When the dcas9-sgRNA complex binds to target sequence, RpoA will recruits 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. | ||
| + | Click BBa_K2996706 for more details, where mRFP is the reporter gene. | ||
Revision as of 15:24, 19 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 (BBa_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 recruits 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.
Click BBa_K2996706 for more details, where mRFP is the reporter gene.
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]