Part:BBa_K4006001

arsC

This part contained a protein coding sequence for arsenate reductase that has been codon optimized for use in the chloroplast of Chlamydomonas reinhardtii. Arsenate reductase catalyzes the reduction of arsenate [As(V)] to arsenite [As(III)]. Using 2 µM arsenate at a substrate, the Kcat of the protein has been recorded as 4.5 min(-1).

We were able to clone this construct into our plasmid and select for transformed E. coli colonies.

Digestion of the miniprepped DNA in the plasmid pASapI with XbaI and BstXI should result in two bands of approximate sizes 4446 bp and 2658 bp as compared to the original plasmid which should have three bands of sizes 4446, 1376, and 800 bp. The top band in each of the columns is undigested DNA, indicating successful cloning of each of the four colonies.

Unfortunately, we were unable to successfully transform this construct into C. reinhardtii and test expression or effect on arsenic uptake in the algae.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]