Part:BBa_M36789

[apFAB46][apFAB687BCD9][NarL][apFAB391][BBa_K216005]

The first component is a strong constitutive promoter (apFAB46 from BioFab database), followed by a bicistronic RBS (apFAB687BCD9 from BioFab database), linked up to production of the NarL gene downstream (P0AF28 from UniProt database). This constitutively promotes NarL, which is one of the regulatory proteins for the napF operon, which encodes for periplasmic nitrate reductase (see Darwin, A., Ziegelhoffer, E., Kiley, P., Stewart, V. 1998. Fnr, NarP, and NarL Regulation of Escherichia coli K-12 napF (Periplasmic Nitrate Reductase) Operon Transcription In Vitro. Journal of Bacteriology, p. 4192-4198). This constitutive production ensures that there will be extra NarL in the cell if the PyeaR promoter diverts all of the naturally produced NarL from other cell functions. A strong terminator with 99% termination efficiency follows (apFAB391 from BioFab database), which is crucial because the constitutive promoter could send a false signal to the PyeaR promoter downstream of the NarL expression region. The last component is the PyeaR promoter (BBa_K216005 from Parts Registry), designed by the 2009 Edinburgh iGEM team. This promoter is regulated by phosphorylated NarL and is responsive to nitrate, nitrite, and nitric oxide in E. coli.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 190
  
• 1000
  COMPATIBLE WITH RFC[1000]