Part:BBa_K1886016
AND GATE-input_ara+IPTG
This is the input part of AND GATE. After being induced by IPTG, plac promoter activates the expression of T7 RNA polymerase which contains two amber mutations (T7 ptag) . After being induced by L-arabinose, pbad promoter activates the expression of a special tRNA (supD), which is used to identify amber mutation. In the wild-type E.coli, amber mutation will inhibit the translation of T7 RNA polymerase. Therefore, only when the two inputs exist together, T7 RNA polymerase can be expressed, and then T7 promoter can be activated to express the downstream gene.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 171
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 174
Illegal BsaI.rc site found at 4155
Characterization
BACKGROUND
Overview
We simplified the three-plasmid system based on our reference by constructing all the segments in one plasmid. And we tested in E.coli MG1655. We used arabinose promoter (PBAD) and lactose promoter (Plac) as inducers and we have verified the corresponding concentration and time.
Principle
This is the input part of AND GATE. After being induced by IPTG, plac promoter activates the expression of T7 RNA polymerase which contains two amber mutations (T7 ptag) . After being induced by L-arabinose, pbad promoter activates the expression of a special tRNA (supD), which is used to identify amber mutation. In the wild-type E.coli, amber mutation will inhibit the translation of T7 RNA polymerase. Therefore, only when the two inputs exist together, T7 RNA polymerase can be expressed, and then T7 promoter can be activated to express the downstream gene.
RESULTS
Gel electrophoretic analysis
stable oscillation in microfluidics chips
| None |