Part:BBa_K3332023

phnD

Subunit of phosphonate ABC transporter, phosphonate binding protein, from S.meliloti 1021.Use BBa_K823004 to construct a new part that can transport glyphosate to cytoplasm.

Biology

        Phn system is a gene cluster for organophosphorus transport and degradation in many microorganisms. Sinorhizobium meliloti 1021 encode ABC transporter by phnEE, phnC, phnD gene, this transporter can transport glyphosate to cytoplasm. The phnC gene encodes ATP-binding protein and the phnD encodes phosphonate binding protein of the ABC transporter.

Usage

We ligased the strong promoter (BBa_J23100) and the parts (RBS-phnE1-RBS-phnE2-RBS-phnC-RBS-phnD-Terminator) on the expression vector pSB1C3 by standard assembly. Then the ligation mixture was transformed into E. coli DH5α & E. coli BL21(DE3), enabled the E. coli to transport glyphosate to cytoplasm at higher efficiency.

Characterization

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 275
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 574
  Illegal AgeI site found at 832
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 177
  Illegal BsaI.rc site found at 511
  Illegal SapI site found at 13