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Part:BBa_K4044011
p119_G4-n11 (Pj23119-based partially modified Gal4 dependant promoter)
The role of this part is genetical regulating systems based on Gal4 as there are no other Gal4 dependant promoters presented in bacteria. When binding to promoter Gal4 domain inhibits transcription activity completely. The promoter contains the sequence of CGG-N11-CCG between GACA-box and TATA-box, which can be recognized by Gal4 protein. Eleven nucleotides in the core location are pj23119 promoter nucleotides. When binding to promoter Gal4 domain inhibits transcription activity completely. This promoter was tested to be active in E. coli and comparable with Anderson promoter pj23119 baseline transcriptional activity. Promoters were tested in construction containing YFP under promoter that is tested with strong RBS used (BBa_J34801). The results of QPAS1-Gal4 inhibition show a significant decrease of fluorescence and result in a close to zero level of transcription.
We tested both base activity of these promoters and the inhibition efficiency of Gal4 with them to get a proper characterisation of these parts. The baseline activity was tested by YFP output with strong RBS used (BBa_J34801). We compared the relative activity of modified promoters with the original J23119 promoter. As a negative control also, original promoter was taken. Thus we get two new promoters dependant on Gal4 that can be used in procaryotic organisms that we consider to be a significant part improvement. Our further step is to try other Anderson promoters to create a library of bacterial Gal4 dependant promoters that there could be more chance to select the best fitting element for different chassies.
We also have shown with a computer simulation that HTH domains of Gal4 binds exactly to CGG-N11-CCG site of the promoter.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 30
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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