Part:BBa_K1031224

Po-B0034-sfGFP-Terminator (DmpR)

For detailed information concerning DmpR and Po promoter, please visit 2013 Peking iGEM Biosensor DmpR

Structure

Po promoter which is activated by DmpR, is σ-54 dependent. It is composed of three regions. The UAS site containing two inverted region for binding is responsible for interaction with DmpR transcriptional factor. The two IHF binding sites allowing IHF to participate enhance transcription efficiency. -24 and -12 region interact with σ-54 factor of RNA polymerase, enabling the formation of open complex. (Fig 1)

Fig 1 Po promoter structure. The UAS of this promoter marked in green is combined of two parts in contrast direction to which DmpR binds. The box with yellow background represents IHF binding sites. The box with pink background represents σ54 binding site with -24 region and -12 region marked in red. The G with right angle represents +1 site.

Sequence and Features

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 210
  

Construction

We constructed a library of RBS (Ribosome Binding Site) of different strength for tuning the expression of reporter sfGFP. K1031223 is a reporter circuit composed of three elements, the inducible promoter Po, RBS B0034, and reporter gene sfGFP with terminator B0015. (Fig 2)

Fig 2 Construction of reporter circuit. The orange arrow represents Po promoter. The green oval stands for RBS B0034. sfGFP coding sequence is shown with dark blue, while terminator B0015 is in dark red.