Part:BBa_K4194014
p(Bla)-RBS-(c-di-GMP riboswitch)-mRFP1-LVA-RBS-TetR-LVA-terminator
The expression of mRFP1 and TetR is regulated by promoter P (Bla) and c-di-GMP riboSwitch. Promoter p (Bla) itself is a moderately strong constitutive promoter, but its downstream c-di-GMP riboswitch enables it to successfully initiate downstream gene expression only when the concentration of c-di-GMP is higher than a certain value, because the c-di-GMP riboswitch can specifically sense the concentration of intracellular c-di-GMP. When the concentration of c-di-GMP increases, it is turned on. The mRFP1 was used as a reporter gene. The Tet-regulated gene expression system was established based on the Tet-resistant operon on the Tn10 transposon of Escherichia coli. TetR has specific affinity with DNA sequences in TetO. TetR binds to two tetracycline manipulation sites (tetO) in p(TetR) with high affinity and inhibits the transcription of downstream genes, which can be relieved by tetracycline. In addition, ssrA-LVA tag was added to the downstream of mRFP1 and TetR in order to allow the protein to be degraded in time.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 838
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 720
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 141
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