Coding

Part:BBa_K5111003

Designed by: Angus Lusty   Group: iGEM24_City-of-London-UK   (2024-09-24)

CDSs for NosR and NosZ

This part contains the coding sequences of the proteins NosR and NosZ. The sequences for the ORFs were taken from P. stutzeri Zobell (ATCC 14405) (https://www.ncbi.nlm.nih.gov/datasets/genome/GCF_015291885.1/), which were subsequently codon optimised for E. coli.

Coded by the gene nosZ, N2OR is a metalloprotein that is found in the periplasm of bacteria and has two distinct copper centres called CuA and CuZ which are both involved in the reduction of nitrous oxide (Zhang et al., 2019). NosR regulates the transcription of nosZ but more importantly acts as an electron donor for N2O reduction (Bennett et al., 2020).

The ability of P. stutzeri Zobell to reduce nitrous oxide in aerobic conditions has been demonstrated, and so we believe this is a promising candidate for aerobic reduction of nitrous oxide (Z. Wang, 2023).

Our part retains the sequence between the two ORFs, which contains the naturally-occurring RBS.


Sources


Zhang, L., Wüst, A., Prasser, B., Müller, C. and Einsle, O. (2019a). Functional assembly of nitrous oxide reductase provides insights into copper site maturation. Proceedings of the National Academy of Sciences, 116(26), pp.12822–12827. doi:https://doi.org/10.1073/pnas.1903819116.

Bennett, S.P., Soriano-Laguna, M.J., Bradley, J.M., Svistunenko, D.A., Richardson, D.J., Gates, A.J. and Le, N.E. (2019). NosL is a dedicated copper chaperone for assembly of the CuZcenter of nitrous oxide reductase. Chemical Science, 10(19), pp.4985–4993. doi:https://doi.org/10.1039/c9sc01053j

Wang, Z., Vishwanathan, N., Kowaliczko, S. and Ishii, S. (2023). Clarifying Microbial Nitrous Oxide Reduction under Aerobic Conditions: Tolerant, Intolerant, and Sensitive. Microbiology Spectrum, [online] 11(2). doi:https://doi.org/10.1128/spectrum.04709-22.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 297
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 255
    Illegal NgoMIV site found at 565
    Illegal NgoMIV site found at 2130
    Illegal NgoMIV site found at 3904
    Illegal AgeI site found at 2336
    Illegal AgeI site found at 2987
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None