Part:BBa_J100295
DB Biosensor + NarX
This construct is built by modifying DB Nitrate Biosensor (J100260), which is built by inserting narG promoter sequence into pClone Red (J119137) through GGA. There are binding sites for activators NarL on the narG promoter. NarX, the sensor kinase, is known to phosphorylate NarL, which is the response regulator, so we inserted the NarX gene into the plasmid replacing the reverse GFP, hoping that will increase NarL binding on the promoter. Also sensor kinase mRNAs are synthesized efficiently but they rapidly degrade. Therefore we inserted a 5' terminal stemloop upstream of the NarX gene to reduce mRNAs degradation by RNase E. In conclusion, we removed the GFP reverse and inserted a complex with a transcription terminator, a promoter, a stemloop, a RBS, and the NarX gene. The goal is that nitrate induces the promoter, then RFP is expressed when nitrate is present. However, this promoter does not induce RFP production when nitrate is present.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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