Difference between revisions of "Part:BBa K3777025"

Latest revision as of 02:56, 22 October 2021

KB2-CtcS-T7(ctcO)-3WJdB-PctcS-sgRNA

Basic biosensor device for Chlortetracycline detection.

Usage and Biology
Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)
When CTR was absent, ctcs would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene. But KB2 will bind to 3WJDB leading to lower fluorescent .
Fig.1 Schematic overview of the genetic circuit.

Reference：Alam Khalid K,Tawiah Kwaku D,Lichte Matthew F,Porciani David,Burke Donald H. A Fluorescent Split Aptamer for Visualizing RNA-RNA Assembly In Vivo.[J]. ACS synthetic biology,2017,6(9)
Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 260
Illegal NheI site found at 283
Illegal NheI site found at 1164
Illegal NotI site found at 832
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 121
Illegal XhoI site found at 841
Illegal XhoI site found at 1305
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 576

@@ Line 8: / Line 8: @@
 <b><font size="3">Usage and Biology</font></b>
 <br>Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)
-<br>When CTR was absent, ctcs would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression.
+<br>When CTR was absent, ctcs would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene. But KB2 will bind to 3WJDB leading to lower fluorescent .
 https://static.igem.org/mediawiki/parts/thumb/d/d9/KB2-CtcS-T7%28ctcO%29-3WJdB-PctcS-sgRNA.PNG/798px-KB2-CtcS-T7%28ctcO%29-3WJdB-PctcS-sgRNA.PNG
 <br>                           Fig.1  Schematic overview of the genetic circuit.