Part:BBa_K1256014

pSB1C3-Pred-ccdB-light sensor

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 [http://2013.igem.org/Team:NCTU_Formosa 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