Part:BBa_K3202046
AraC-Pc-pBAD-LacI-B0034-msfGFP-T500-T1/TE-MicCsRNA-sRNA Binding Site-PA1/O4
This composite part is a modified version of part BBa_K3202044; RBS is replaced with B0034 In our study, the initial design contained a RBS that included a terminator of former coding sequence, and therefore it failed. In this design, we replaced the RBS by an ordinary one, B0034, to check the functionality of other components in the sequence. 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Illegal SapI.rc site found at 2361
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