Difference between revisions of "Part:BBa K337049"

 
 
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<partinfo>BBa_K337049 short</partinfo>
 
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[[Image:PSMB_miMeasure.png|thumb|250px|right|pSMB_miMeasure: basic scheme]]
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pSMB_miMeasure is the standard fluorescent-based microRNA binding site characterization vector. It is composed of a mirrored construct setup. Two fluorescent proteins EGFP and EBFP2 are driven by a bidirectional CMV promoter to ensure the same expression strength at both sides. The two fluorescent proteins are destabilized by fusion to a MODC degradation domain which leads to a protein half life of two hours. The shortened half life of the destabilized fluorescent proteins enables for time-laps experiments in order to analyze binding site properties in a  dynamic microRNA environment. Furthermore the plasmid contains a BBb_2 (RFC12, RFC41) standard site behind the EGFP to enable user friendly exchange of binding site of choice. BamHI and HindIII sites behind the EBFP2 enable for cloning in a reference binding site. The plasmid further contains an ampicillin resistance for selection in bacteria. As the complete Plasmid has been synthesized by Geneart, it is in the pMA backbone (BB-2, RFC12).
  
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microRNA binding site measurment plasmid
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Latest revision as of 17:56, 29 October 2010

pSMB_miMeasure

pSMB_miMeasure: basic scheme


pSMB_miMeasure is the standard fluorescent-based microRNA binding site characterization vector. It is composed of a mirrored construct setup. Two fluorescent proteins EGFP and EBFP2 are driven by a bidirectional CMV promoter to ensure the same expression strength at both sides. The two fluorescent proteins are destabilized by fusion to a MODC degradation domain which leads to a protein half life of two hours. The shortened half life of the destabilized fluorescent proteins enables for time-laps experiments in order to analyze binding site properties in a dynamic microRNA environment. Furthermore the plasmid contains a BBb_2 (RFC12, RFC41) standard site behind the EGFP to enable user friendly exchange of binding site of choice. BamHI and HindIII sites behind the EBFP2 enable for cloning in a reference binding site. The plasmid further contains an ampicillin resistance for selection in bacteria. As the complete Plasmid has been synthesized by Geneart, it is in the pMA backbone (BB-2, RFC12).





Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2643
    Illegal SpeI site found at 2657
    Illegal PstI site found at 2680
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal prefix found in sequence at 2643
    Illegal NheI site found at 2665
    Illegal PstI site found at 2680
    Illegal NotI site found at 2673
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2643
    Illegal BamHI site found at 257
    Illegal XhoI site found at 2913
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2643
    Illegal SpeI site found at 2657
    Illegal PstI site found at 2680
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2643
    Illegal SpeI site found at 2657
    Illegal PstI site found at 2680
  • 1000
    COMPATIBLE WITH RFC[1000]