Part:BBa_K2207997

2,4-DAPG PhlABCD Cluster with T7 promoter and Double Terminator

We successfully cloned the phlABCD via colony PCR from Pseudomonas fluorescens 2P24. We have submitted it as a basic part 2,4-DAPG PhlABCD Cluster(BBa_K2207998),and then we put BBa_K2207998 under the control of a strong promoter which is present in the plasmid backbone we used. This backbone(BBa_K525998) was obtained from the part registry.And we use Double Terminator(BBa_B0015) to stop the transcription.

Upon transformation of this biobrick into BL21 E. coli cells and induced with IPTG，our engineered bacteria will synthesize DAPG.We utilize the High-performance liquid chromatography (HPLC) method to detect the biosynthesized DAPG. Luckily, we finally detected the DAPG from E.coli cells as Fig.1. shows.

Fig.1. Result of DAPG detecting by HPLC.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 672
  Illegal XhoI site found at 1538
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 85
  Illegal NgoMIV site found at 372
  Illegal AgeI site found at 1847
  Illegal AgeI site found at 1875
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3725