Part:BBa_K2207014

PhlAC enzyme

DAPG is initially found in specific strain of Pseudomonas fluorescens. And it has been demonstrated that a cluster named phl responsible for the biosynthesis of DAPG, which contains eight genes, from phlA to phlH. Further studies shows that phlABCD are the DAPG synthesis genes.We successfully cloned the phlABCD via colony PCR from Pseudomonas fluorescens 2P24. In this part, we inserted two of them, phlA and phlC, linked with F2A sequence,which can convert MAPG to DAPG together with phlB.

We successfully inserted this part into vector pKD1 and transformed it into T. atroviride, and used fluorescence microscope to detect the hyphae of our engineered T. atroviride. To our joy, we successfully saw the green fluorescence as Fig.1 shows, which means phlAC both expressed in our engineered T. atroviride.

Fig.1 Result of fluorescence detecting.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 4607
  Illegal PstI site found at 776
  Illegal PstI site found at 4619
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 776
  Illegal PstI site found at 4619
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1494
  Illegal XhoI site found at 2168
  Illegal XhoI site found at 3079
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 4607
  Illegal PstI site found at 776
  Illegal PstI site found at 4619
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 4607
  Illegal PstI site found at 776
  Illegal PstI site found at 4619
  Illegal NgoMIV site found at 1298
  Illegal NgoMIV site found at 1581
  Illegal NgoMIV site found at 1868
  Illegal AgeI site found at 3388
  Illegal AgeI site found at 3416
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 486
  Illegal SapI site found at 168