Difference between revisions of "Part:BBa K3332022"

Revision as of 18:25, 27 October 2020

phnC

Subunit of phosphonate ABC transporter, ATP 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 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.

Usage

We ligased the coding part J23100-B0034-phnE1-B0034-phnE2 (BBa_K3332067) and the part RBS-phnC-RBS-phnD 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
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 112
Illegal NgoMIV site found at 181
Illegal NgoMIV site found at 420
Illegal NgoMIV site found at 424
Illegal NgoMIV site found at 805
Illegal NgoMIV site found at 829
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 192
Illegal SapI.rc site found at 358

@@ Line 10: / Line 10: @@
 ===Usage===
-&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We ligased the coding sequences of phnE1 and phnE2 (RBS-phnE1-RBS-phnE2) and the part J23100-RBS-phnE1-RBS-phnE2-J23100-RBS-phnJ-Terminator(<partinfo>BBa_K3332097</partinfo>) 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.
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We ligased the coding part J23100-B0034-phnE1-B0034-phnE2 (<partinfo>BBa_K3332067</partinfo>) and the part RBS-phnC-RBS-phnD 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===