Translational_Unit

Part:BBa_K4773000:Design

Designed by: Yan Pan   Group: iGEM23_UESTC-China   (2023-09-27)


RBS-NicA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 123
    Illegal BglII site found at 364
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The expression of NicA may be regulated by an inducible promoter, such as BBa_I13453 (pLacIq), to control the timing and level of nicotine degradation.

The expression of NicA may affect the growth and viability of the bacterial cells, due to the production of pseudooxynicotine and other toxic intermediates. Therefore, it may be necessary to optimize the culture conditions and/or provide detoxification mechanisms for the bacterial cells.


Source

Source: This part is derived from the genomic sequence of Pseudomonas putida S16, a nicotine-degrading bacterium isolated from tobacco waste. And was cloned into the pUSP vector with the BBa_B0034 RBS.

References

Tang H, Wang L, Wang W, Yu H, Zhang K, Yao Y, Xu P. Systematic unraveling of the unsolved pathway of nicotine degradation in Pseudomonas. PLoS Genet. 2013 Oct;9(10):e1003923. doi: 10.1371/journal.pgen.1003923. Epub 2013 Oct 31. PMID: 24204323; PMCID: PMC38143101

Wang L, Tang H, Wang S, Liu R, Liu M, Zhang Y, Chen T, He X, Li S, Xu P. Novel nicotine oxidoreductase-encoding gene involved in nicotine degradation by Pseudomonas putida strain S16. Appl Environ Microbiol. 2012 Jan;78(2):364-72. doi: 10.1128/AEM.06262-11. Epub 2011 Nov 4. PMID: 22057864; PMCID: PMC32556472