Generator

Part:BBa_J33201

Designed by: Chris French   Group: iGEM06_Edinburgh   (2006-10-13)
Revision as of 02:13, 19 October 2019 by Lavenderxc (Talk | contribs)


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 255
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



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. 

  T--TJU_China--1-8.png

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. 

  T--TJU_China--1-9.png

Figure2. The result of nucleic acid gel electrophoresis of Ars Promoter after PCR. lane M, Marker. Lane 1-4, Ars Promoter. 

  T--TJU_China--1-10.png

Figure 3. The result of nucleic acid gel electrophoresis of ArsR Protein after PCR. Lane M, Marker. Line1, ArsR Protein.

Characterization

Worldshaper-Wuhan 2019's Characterization

Here, we used J33201 part to construct a biosensor pSB1C3-pArsR-RBS-GFP(K3153000) to detect the arsenite(As3+) in water, and to add a contribution to this well-characterized part.

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