Part:BBa_K1913024
Synthetic ptet-FixK2 hybrid promoter with RBS
Synthetic ptet-FixK2 hybrid promoter is a promoter wich can not only be activated by light sensors YF1-FixJ composite but also inhibited by the TetR repressor. The sequence of the upper control element is from wild type FixK2 promoter on the genomic sequence of Bradyrhizobium japonicum, which contains two typical FixJ boxes. The core element region is replaced by the core element region of BBa_J23106. Additional tet operators are inserted into the core element region of promoter as second control elements which result in transcription repression. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 118
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 27
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
In order to test the hybrid promoters, we constructed five composite parts with mRFP gene as reporter (BBa_K19103027, BBa_K19103028, BBa_K19103029, BBa_K19103030, BBa_K19103031). These composite parts were co-transformed with the light sensor part (BBa_K19103034) into E.coli strain BL21, cultured under dark for 24h and their fluorescence was tested. We included as controls cells that only contained the Fixk2 composite parts. The results (Fig.1) illustrated that hybrid promoter BBa_K1913024 has even less fluorescence value compared to control, which suggested that this promoter does not response for being induces by FixJ.
Figure 1. Ratio of fluorescence value and absorbance of Fixk2 composite. Fixk2 composite and control was cultured under dark condition over 24h. Emission and excitation wavelength of mRFP are 607 and 584 nm respectively.
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