Part:BBa_K346000
RBS(B0032)+T3 DNA-directed RNA polymerase
T3 polymerase can activate the expression of different T3 promoters in T3 phage. Since during different time the phage need different expression level of different genes, there must be T3 promoters of variable sensitivity. In our experiment, we constructed 14 different T3 promoter by primer annealing and by standard digestion to put it into standard plasmid. We then add GFP downstream to function as a reporter. By transforming another plasmid contains T7 promoter+RBS+T3 polymerase, we were able to detect the expression level of different promoters. We finally constructed 9 promoters with variable sensitivity, and put them in order according to the GFP expression level. We found that among them the Phi9 T3 promoter has a medium expression level and was more robust in the system, so we chose this one to be used in our kit. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2585
//function/regulation/transcriptional
//rnap/bacteriophage/t3
| biology | Orthogonal to T7 polymerase/T7 promoter pair |
| family | Phage and mitochondrial RNA polymerase family |
| function | DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. |
| n/a | RBS(B0032)+T3 DNA-directed RNA polymerase |
| negative_regulators | Bacterial RNA polymerase inhibitor from T3 phage |
| origin | Enterobacteria phage T3 (Bacteriophage T3) |
| rbs | Medium |
| uniprot | P07659 |