Composite

Part:BBa_K3739067

Designed by: Shichen Geng   Group: iGEM21_XMU-China   (2021-09-09)


J23100-B0034-phnC-B0034-phnD-J23100-B0034-phnE1-B0034-phnE2

Phn system is a gene cluster for organophosphorus transportation and degradation in many microorganisms. Sinorhizobium meliloti 1021 encodes 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.

Biology

Phn system is a gene cluster for organophosphorus transport and degradation in many microorganisms. Sinorhizobium meliloti 1021 use PhnE1 and PhnE2 to construct the permease protein of ABC transporter, which includes PhnE1, PhnE2, PhnC and PhnD proteins, this transporter can transport glyphosate to cytoplasm. The phnEE gene encodes permease protein which can transport glyphosate to cytoplasm. The phnC gene encodes ATP-binding protein and the phnD encodes phosphonate binding protein of the ABC transporter.

Usage

To improve the efficiency of this system in E. coli, we constructed the composite part BBa_J23100-BBa_B0034-BBa_K3332022-BBa_B0034-BBa_K3332023-BBa_K3332067 on the expression vector pSB1C3, the constructed plasmid was transformed into E. coli BL21(DE3), then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1848
    Illegal NheI site found at 1871
    Illegal NheI site found at 2898
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1202
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 173
    Illegal NgoMIV site found at 242
    Illegal NgoMIV site found at 481
    Illegal NgoMIV site found at 485
    Illegal NgoMIV site found at 866
    Illegal NgoMIV site found at 890
    Illegal NgoMIV site found at 1501
    Illegal AgeI site found at 1759
    Illegal AgeI site found at 4050
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 253
    Illegal BsaI site found at 1104
    Illegal BsaI.rc site found at 1438
    Illegal SapI site found at 940
    Illegal SapI.rc site found at 419


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Categories
Parameters
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