Part:BBa_K2150027
pCon+RBS+tetR+DT+pT7+pTet+RBS+tetX-GFP(fusion protein)+RBS+T7RNAP+DT
This part is a complete regulatory device sensitive to tetracycline. A fusion protein(referred to as tetX-GFP) of tetX(a tetracycline-degrading enzyme) and GFP combined with a 3*GGGGS linker is used to report the expression level of tetX. The first promoter(BBa_J23119) is constitutively on and can express tetR, a protein that can repress the downstream promoter pTet in the absence of tetracycline(Tc). As a result, no tetX-GFP is produced and no fluorescence can be detected. In the presence of Tc, however, tetR can be repressed by Tc and repression over pTet can be removed, resulting in the expression of tetX-GFP and T7RNAP. Under such circumstances, activity of tetX can be reported by green fluorescence and this activity can be amplified by T7RNAP-pT7 system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1561
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2858
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