Part:BBa_K4192132

Plac-RBS-yf1(N37C)-RBS-fixJ-PfixK2-RBS-mcherry

The gene yf1, fixJ and PfixK2 make up the system of blue light regulation. Under dark conditions, phosphate groups are transferred from Yf1 protein to FixJ protein, and the phosphorylated FixJ protein activates the PfixK2 promoter, which in turn activates downstream gene expression. Under light induction, the phosphorylation of Fixj protein was blocked and the expression of genes regulated by the PfixK2 promoter was suppressed.The mCherry protein emits red fluorescence to characterize the efficiency of control elements in regulating gene expression. We mutated position 37 N of YF1 to C to improve its response to blue light and better regulate gene expression.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 210
  Illegal SpeI site found at 1361
  Illegal SpeI site found at 2257
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 210
  Illegal SpeI site found at 1361
  Illegal SpeI site found at 2257
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 210
  Illegal SpeI site found at 1361
  Illegal SpeI site found at 2257
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 210
  Illegal SpeI site found at 1361
  Illegal SpeI site found at 2257
  Illegal NgoMIV site found at 764
  Illegal NgoMIV site found at 836
  Illegal NgoMIV site found at 926
  Illegal NgoMIV site found at 944
  Illegal NgoMIV site found at 1450
  Illegal NgoMIV site found at 1743
  Illegal NgoMIV site found at 1837
  Illegal AgeI site found at 478
  Illegal AgeI site found at 1618
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1507
  Illegal BsaI.rc site found at 377