Difference between revisions of "Part:BBa K3202068"

Revision as of 06:58, 19 October 2019

AraC-Pc-pBAD-RBS1-Colicin E2-T500-T1/TE-MicCsRNA-PA1/O4

This composite part is designed to insert the toxin Colicin E2 into the bistable system.

Presenting with BBa_K3202066-BBa_K3202067, this part is to test the function of the toxin within our bistable system.The double bistable system combines small RNA-mediated inhibition of translation with the translational coupling of a repressor to the gene of interest. In our system, we have transcriptional repressors tetR expressed through the inducible promoter but translationally inhibited by the MicC sRNA1 respectively. Since RBS2 and RBS3 is directly inhibited by sRNA while they initiates the transcription of the two repressors, the gene of interest is translationally coupled to the repressor gene while three promoters which produces the inhibitory sRNA are targeted by their corresponding repressors. Therefore, repressors and sRNA are mutually inhibitory. Within one single layer of the bistable system, the transcriptional factor is turned on, pBAD initiates the expression of tetR, which raises the concentration of tetR and thereby exerts greater inhibition on PR1 and therefore decreases the concentration of sRNA1. As sRNA1 is inhibited, the inhibitory effect it exerts on tetR accordingly decreases; therefore, the recombinase is expressed through the first layer, and vice versa. Nevertheless, given that the expression of tetR inhibits PR1, even if the recombinase is expressed PR1 can be flipped over but cannot express the second layer.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1144
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 979
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 961

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3202068 short</partinfo>
-This composite part is designed to insert the toxin colicin into the bistable system.
+This composite part is designed to insert the toxin Colicin E2 into the bistable system.
 Presenting with BBa_K3202066-BBa_K3202067, this part is to test the function of the toxin within our bistable system.The double bistable system combines small RNA-mediated inhibition of translation with the translational coupling of a repressor to the gene of interest. In our system, we have transcriptional repressors tetR expressed through the inducible promoter but translationally inhibited by the MicC sRNA1 respectively. Since RBS2 and RBS3 is directly inhibited by sRNA while they initiates the transcription of the two repressors, the gene of interest is translationally coupled to the repressor gene while three promoters which produces the inhibitory sRNA are targeted by their corresponding repressors.  Therefore, repressors and sRNA are mutually inhibitory. Within one single layer of the bistable system, the transcriptional factor is turned on, pBAD initiates the expression of tetR, which raises the concentration of tetR and thereby exerts greater inhibition on PR1 and therefore decreases the concentration of sRNA1. As sRNA1 is inhibited, the inhibitory effect it exerts on tetR accordingly decreases; therefore, the recombinase is expressed through the first layer, and vice versa. Nevertheless, given that the expression of tetR inhibits PR1, even if the recombinase is expressed PR1 can be flipped over but cannot express the second layer.