Plasmid

Part:BBa_K1256014

Designed by: Pei-Hong Chen   Group: iGEM14_Mingdao   (2014-06-10)

pSB1C3-Pred-ccdB-light sensor

Mingdao pSB1C3-Pred-ccdB-light sensor.png

Gene expression of ccdB (BBa_K145151), a lethal gene, is regulated by the light. The light-regulated system is composed of light sensor (BBa_K1017101) and light-regulated promoter (BBa_K1017301) designed and constructed by NCTU-Formosa in 2013. Light sensor was first amplified by PCR, cut by EcoRI and PstI, and ligated to pSB1C3-Plac-NB-Cm-MN cut by MfeI and NsiI. Then, light-regulated promoter was amplified by PCR, cut by EcoRI and NheI, and ligated to the resulting vector cut by EcoRI and NheI, which maintains the standard BioBrick assembly rule as well as the flexibility of exchanging RFP cds with NheI and BamHI. The ccdB was amplified by PCR using the forward primer with RBS and assembled to the vector with NheI and BamHI.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 6415
    Illegal suffix found in sequence at 2092
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6415
    Illegal NheI site found at 1619
    Illegal SpeI site found at 2093
    Illegal PstI site found at 2107
    Illegal NotI site found at 2100
    Illegal NotI site found at 6421
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6415
    Illegal BglII site found at 1282
    Illegal BglII site found at 3244
    Illegal BamHI site found at 1949
    Illegal XhoI site found at 3124
    Illegal XhoI site found at 5137
    Illegal XhoI site found at 6291
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 6415
    Illegal suffix found in sequence at 2093
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 6415
    Illegal XbaI site found at 6430
    Illegal SpeI site found at 2093
    Illegal PstI site found at 2107
    Illegal NgoMIV site found at 1296
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1858


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Categories
Parameters
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