Composite

Part:BBa_K593009

Designed by: Ceren Seref   Group: iGEM11_METU-Ankara   (2011-09-21)


ROSE regulated kill switch

This composite part consists of promoter, RNA thermometer, GFP and lysis casette. The upstream part of this composite is T7 promoter (BBa_I712074) which is strong promoter from T7 bacteriophage. For strong transcription T7 promoter expression system is chosen. In downstream of promoter there is RNA thermometer(ROSE) coding region which determines the transcription of our lysis composite part. RNA thermometer (BBa_K115001) is temperature sensitive that at 42C translation initiates. That means, up to 42 C RNA thermometer is in dimer form and at 42C it becomes linear. In downstream of RNA thermometer, green fluorescence protein(BBa_E0040) is ligated. This ligation is done to measure expression level and to comment on RNA thermometer efficiency. The last ligate to this composite is bacteriophage 21 lysis casette (BBa_K124003). This lysis casette induces lysis in E.coli. This gene cassette consists of the S protein (Holin), R protein (Endolysin), and Rz protein. When translation initiates at 42 C then the following coding gene begins to produce the proteins which lyse the cell.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 884
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 884
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 884
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 884
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 884
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1633
    Illegal BsaI.rc site found at 800


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Categories
Parameters
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