E. coli chromosomal ars promoter with arsR repressor gene
This part consists of the promoter of the E. coli JM109 chromosomal arsenic detoxification operon (ars operon), including the ArsR repressor binding site and the arsR gene encoding the arsR repressor protein, together with its ribosome binding site. Addition of any other genes to the 3' end of this part will result in their expression being dependent on the presence of sodium arsenate or sodium arsenite. Arsenite or arsenite anion binds to the repressor protein ArsR, resulting in inability to repress the promoter. Based on our experiments, a concentration of 1 micromolar sodium arsenate in LB is sufficient for essentially full expression, though this will vary according to conditions.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BglII site found at 255
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC
Supplementary material offered by other groups
The following is from TJU-China 2018. We used the part:Bba-J33201 saved in plate3 and wanted to build a new arsenic induction loop through it. Therefore, we first added 10μlddH2O to the corresponding hole, and then took PCR was performed at 1μl, and the corresponding DNA fragment was obtained.
Figure1. The result of nucleic acid gel electrophoresis of Bba-J33201 after PCR. Lane M, Marker. Lane 1-6，Bba-J33201
Then we performed PCR on the promoter fragment and ArsR fragment in this fragment, and the results are as follows.
Figure2. The result of nucleic acid gel electrophoresis of Ars Promoter after PCR. lane M, Marker. Lane 1-4, Ars Promoter.
Figure 3. The result of nucleic acid gel electrophoresis of ArsR Protein after PCR. Lane M, Marker. Line1, ArsR Protein.