Part:BBa_K5111002

ComR CDS in a composite part

This part contains the coding sequence for ComR (BBa_K5111001) (also known as Ycfq) in a composite part containing a promoter, ribosome binding site, and terminator. This sequence is designed to synthesise the ComR protein at a high level (M. Mermod et al., 2017).

ComR downregulates the expression of a separate protein, ComC, involved in decreasing membrane permeability to copper ions. Therefore, as ComR is upregulated, ComC is downregulated, and copper levels in the periplasm and cytoplasm rise.

The part is designed to increase copper bioavailability for nitrous oxide reduction, but the system can theoretically be applied to other areas. Many of the copper-uptake systems already on the registry focus on binding copper for bioremediation, rather than increasing its bioavailability in the organism. Therefore, our ComR system presents an opportunity for other enzymes with copper cofactors, like Cytochrome c oxidase (F. Fontanesi et al., 2008) and superoxide dismutase-1 (K. Sea et al., 2015).

The promoter is used BBa_J23106, a strong promoter used to maximise expression of ComR.

Sources

Mermod M, Magnani D, Solioz M, Stoyanov JV. The copper-inducible ComR (YcfQ) repressor regulates expression of ComC (YcfR), which affects copper permeability of the outer membrane of Escherichia coli. Biometals. 2012 Feb;25(1):33-43. doi: 10.1007/s10534-011-9510-x. Epub 2011 Nov 17. PMID: 22089859.

Fontanesi F, Soto IC, Barrientos A. Cytochrome c oxidase biogenesis: new levels of regulation. IUBMB Life. 2008 Sep;60(9):557-68. doi: 10.1002/iub.86. PMID: 18465791; PMCID: PMC2630494.

Sea K, Sohn SH, Durazo A, Sheng Y, Shaw BF, Cao X, Taylor AB, Whitson LJ, Holloway SP, Hart PJ, Cabelli DE, Gralla EB, Valentine JS. Insights into the role of the unusual disulfide bond in copper-zinc superoxide dismutase. J Biol Chem. 2015 Jan 23;290(4):2405-18. doi: 10.1074/jbc.M114.588798. Epub 2014 Nov 28. PMID: 25433341; PMCID: PMC4303690.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]