Part:BBa_J100293
B Biosensor + NarX
This construct is built by modifying B Nitrate Biosensor (J100258), which is built by inserting a modified 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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1134
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1134
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1533
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1134
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1134
Illegal AgeI site found at 3069
Illegal AgeI site found at 3181 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 627
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