Part:BBa_K4182009
A circuit for efficient exopolysaccharide synthesis
we selected the E.coli-pgmA+E.coli-GalU gene (later referred to as EE gene) with the best EPS expression with the LacI manipulator to form plasmid IV where the GalU gene and the pgmA gene set from E. coli were synthesized into the EE gene, In specific experiments we obtained the EE gene, LacI gene in separate isolation and extraction Figure In our specific experiments, we isolated and extracted EE gene, LacI gene pSB1K3 plasmid vector and recovered them, and then performed GoldenGate ligation by using Bsa I enzyme cleavage site. Because the plasmid copy number was not high, we were unable to obtain the linker. So we changed to high copy pSEVA341 plasmid and re-linked it to obtain new plasmid IV and successfully constructed it
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 4724
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4724
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4724
Illegal BglII site found at 4733
Illegal BglII site found at 7225 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 4724
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 4724
Illegal NgoMIV site found at 1103
Illegal NgoMIV site found at 4610
Illegal AgeI site found at 6785
Illegal AgeI site found at 6951 - 1000COMPATIBLE WITH RFC[1000]
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