Difference between revisions of "Part:BBa K2996704"

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 52
Illegal NheI site found at 75
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 39
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2996704 short</partinfo>
-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.
+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.)
+<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.
-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.
+Check BBa_K2996706 for more details.