Regulatory

Part:BBa_M33001:Design

Designed by: Daniel Bui and Aaditya Shidham   Group: Stanford BIOE44 - S010   (2010-06-01)

Nar operon promoter (narGp) => PoPs


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]


Design Notes

The sequence was checked for restriction digest sites to ensure compliance with Assembly Standard 10. Sequence does not contain an RBS or the narG CDS.

No RBS (ribosome binding sites) could be included in the promoter region, which is why the sequence stops 12 bp before the CDS. This is because the presence of an RBS goes beyond the scope of the promoter device and would constrain a user of the piece to one RBS for the reporter gene.

Source

Part was synthesized based on the promoter of the nar operon naturally present in E. coli as part of nitrate respiration. It was directly copied from the EcoGene DNA database (ecogene.org). The selection was from 156 bp upstream of the start of narG CDS (coding sequence) to 12 bp upstream of the start of CDS.

Works Consulted

1. DeAngelis, K. M., P. S. Ji, M. K. Firestone, and S. E. Lindow. 2005. Two novel bacterial biosensors for detection of nitrate availability in the rhizosphere. Appl. Environ. Microbiol. 71:8537-8547

2. Hartig, Elizabeth. "Nitrate and Nitrite Control of Respiratory Nitrate Reduction in Denitrifying Pseudomonas stutzeri by a Two-Component Regulatory System Homologous to NarXL of Escherichia coli." Journal of Bacteriology 181.12 (1999): 3658–3665. Web.

3. Jayaraman, P. S., T. C. Peakman, S. J. Busby, R. V. Quincey, and J. A. Cole. 1987. "Location and sequence of the promoter of the gene for the NADH-dependent nitrite reductase of Escherichia coli and its regulation by oxygen, the Fnr protein and nitrite." J. Mol. Biol. 196:781-788.

4. Li, S., Rabi, T., & DeMoss, J. A. (1985)."Delineation of two distinct regulatory domains in the 5'region of the nar operon of Escherichia coli." Journal of bacteriology, 164(1), 25.

5. Li, Songfeng, Delineation of Two Distinct Regulatory Domains in the 5' Region of the nar Operon of Escherichia coli. Journal of Bacteriology 164.1 (1985): 25-32.