Difference between revisions of "Part:BBa K1031114"
Line 33: Line 33:
  
 
<html>
 
<html>
−
<img src="https://static.igem.org/mediawiki/igem.org/7/77/Peking2013_part_32-Po-CapR.png", width=500px; />
+
<img src="https://static.igem.org/mediawiki/igem.org/7/77/Peking2013_part_32-Po-CapR.png", width=400px; />
 
</html>
 
</html>
  

Revision as of 08:04, 25 September 2013

Po-B0032-sfGFP (CapR)


Structure

Po promoter which is activated by CapR, is σ-54 dependent. It is composed of three regions. The two inverted UAS sites are responsible for binding of CapR transcriptional factor. The two IHF binding sites allowing IHF to participate enhance transcription efficiency. -24 and -12 region interact with sigma-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 208


Construction

We constructed a library of RBS (Ribosome Binding Site) of different strength for tuning the expression of reporter sfGFP. K1031114 is a reporter circuit composed of three elements, the inducible promoter Po, RBS B0032[1], and reporter gene sfGFP. (Fig 2)

Fig 2 Construction of reporter circuit. The orange arrow represents Po promoter. The green oval stands for RBS B0032. sfGFP coding sequence is shown with dark blue, while terminator B0015[2] is in dark red.