Part:BBa_K2207015

PhlBD 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, phlB and phlD, linked with F2A sequence.

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 phlBD 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 1582
  Illegal XbaI site found at 3794
  Illegal PstI site found at 776
  Illegal PstI site found at 3806
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 776
  Illegal PstI site found at 3806
  
• 21
  INCOMPATIBLE WITH RFC[21]

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

  Illegal XbaI site found at 1582
  Illegal XbaI site found at 3794
  Illegal PstI site found at 776
  Illegal PstI site found at 3806
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 1582
  Illegal XbaI site found at 3794
  Illegal PstI site found at 776
  Illegal PstI site found at 3806
  Illegal NgoMIV site found at 1298
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 486
  Illegal BsaI.rc site found at 2279
  Illegal SapI site found at 168