Part:BBa_J33203

E. coli ars promoter, arsR gene and lacZ

This biobrick contains the promoter region of the Escherichia coli chromosomal arsenic detoxification opern, including the arsR gene encoding the ArsR repressor protein required for inducibility of this promoter. This is fused to a lacZ' minigene encoding the N-terminal 77 amino acid residues of LacZ, sufficient to complement the lacZ-delta-M15 mutation. When introduced into an E. coli host strain carrying the lacZ-delta-M15 mutation, this construct results in lacZ expression in the presence of arsenate or arsenite anion. LacZ expression can be detected in various ways. This construct was designed for the detection of arsenate and arsenite in water samples.

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]

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Contribution

Group: NCTU_Formosa 2017

Author: Yi-Ting Lin, Huei-Yu Yeh, Sheng-Ping Chu

New improved part Improvement

Summary

BBa_J33201 is a promoter containing the E. coli JM109 chromosomal arsenic detoxification operon (ars operon). Both 2013 Buenos Aires iGEM team and 2006 Edinburgh iGEM team used it to detect arsenic ions in the water. Buenos Aires team ligased it with GFP and Edinburgh team ligased it with LacZ α. The two teams also submitted biobricks, BBa_K1106004 and BBa_J33203. We wanted to know whether these two biobricks have more application, such as detecting the arsenic pollution in Chinese medicine. We also wanted to know whether they can detect other metal ions. Several experiments were done to evaluate the function of this promoter.