Coding

Part:BBa_K2363000

Designed by: Mary Elizabeth Adler   Group: iGEM17_iTesla-SoundBio   (2017-10-25)
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PCBA5 from Dehalococcoides mccartyi

This part contains a coding site for the PCBA5 gene, extracted from the organism d.mycarti. The resultant enzyme is a reductive dehalogenase and is able to degrade PCB toxins by stripping away chlorine groups from the overall structure. Three gene strains of this nature were identified by in 2014 by Dr. Shanquan Wang et. all, CG1, CG4, and CG5 (renamed PCBA1, PCBA4, and PCBA5). We used NCBI to acquire the sequences of each gene (accession number CP006951.1), then codon optimized and synthesized each one. This is the coding region only (1442 base pairs) and does not contain any promoter or RBS regions. We did, however, add a start and stop codon. We found a high confidence interval between the PCBA sequences and pceA (a well-characterized reductive dehalogenase), leading us to believe that they have the same interaction with the cofactor norpseudo B12 (Krautler 2003).


Wang S, Chng KR, Wilm A, et al. Genomic characterization of three unique Dehalococcoides that respire on persistent polychlorinated biphenyls. Proceedings of the National Academy of Sciences of the United States of America. 2014;111(33):12103-12108. doi:10.1073/pnas.1404845111.

Krautler, Bernhard. The Cofactor of Tetrachloroethene Reductive Dehalogenase of Dehalospirillim multivorans Is Norpseudo-B12, a New Type of a Natural Corrinoid. Helvetica Chimica Acta. 2003;vol86:3698. 10/31/17


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]


[edit]
Categories
Parameters
None