Part:BBa_K2570019
tetR+GFP+mazF+PLtetO1+RFP
The Tet-Off system which turns expression of its controlling genes OFF by the addition of tetracycline and its derivative is often used for this purpose and has been applied to several organisms.
The figure shows a strategy for metabolic state switching module using a synthetic metabolic regulator.
AraC repressor inhibits transcription from the PBAD promoter. When transcription by the PBAD promoter is induced with arabinose addition, expression of tetR and GFP is promoted and the expression of mCherry under the PLtetO1 promoter is inhibited by the TetR repressor. Without arabinose addition, expression of tetR and GFP under the PBAD promoter is inhibited by the araC repressor and the expression of mCherry under the PLtetO1 promoter is promoted.
For the 2-PE part, we developed a switch which realize the transform from producing mode to suicide mode. TetO & TetR are common logic elements used in synthetic biology, linking with 2-PE and mazf. Without the induction of arabinose, TetR and mazf won’t be expressed, and production of 2-PE maintained at a normal level. anhydrotetracycline (ATc) is a tetracycline analog, showing increased (30x) affinity for tet repressor. 42 ng/mL gives induction at half maximum according to an in vitro assay. If the user would like to stop the 2-PE production, adding tiny concentration of arabinose will be able to achieve. TetR will be expressed to inhibit TetO and mazf caused cell death.We currently use GFP and RFP to characterize the expression of our circuits. And we’re also trying to get further data results. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2590
Illegal AgeI site found at 2702 - 1000COMPATIBLE WITH RFC[1000]
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