Reporter

Part:BBa_K352011

Designed by: Ş.Serap Memik   Group: iGEM10_METU_Turkey   (2010-10-16)

pCooM Promoter coupled with RBS+RFP+Double terminator

pCooM Promoter coupled with RBS, RFP,double terminator


This component is used to measure high levels of carbon monoxide in the environment. Construction of this device was performed by utilizing strong CO sensitive promoter which is pCooM promoter from Rhodospirillum rubrum. This device is only active when there is CooA transcriptional activator and Carbon monoxide gas in the environment. When there is CO in the environment, the device produces red fluorescent protein in response


PcooF and PcooM, the two CooA-regulated R. rubrum promoters, contain 2-fold symmetric DNA sequences which are CooA-binding sites and are similar to the CRP consensus sequence. This similarity corralates with the tantamount helix-turn-helix motifs CooA and CRP. The CooA binding sites lie at the 243.5 and 238.5 positions relative to the transcription start sites in PcooF and PcooM, respectively, overlapping with the 235 region. This overlap suggests that both CooA-regulated promoters are analogous to class II CRP-dependent promoters. We chose PcooF for this study because it is the stronger promoter based on the amount of primer extension product and level of coo-encoded proteins synthesized in vivo.




Imaging with confocal laser scanning microscopy

w7

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 647
    Illegal AgeI site found at 759
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds/ligand
//chassis/prokaryote/ecoli
//classic/reporter
Parameters
emission510 nm
excitation395 nm
ligandsCO bound cooA
n/apCooM Promoter coupled with RBS+RFP+Double terminator
outputRFP