Description

pAUR135 is a shuttle vector including DNA sequence to remove the vector region containing selectable marker AUR1-C from transformations of Saccharomyces cerevisiae by this vector. Selection of marker-removed clones is easy and efficient, and so pAUR135 vector is available repeatedly for transformation.
The vector contains AUR1-C gene that confers Aureobasidin A (AbA)-resistance on yeast cells and GIN11M86 DNA under the control of GAL10 promoter. Overexpression of GIN11M86 causes growth inhibition of host cells. Most of AbA-resistant transformants by pAUR135 vector, when shifted onto galactose medium which induce overexpression of GIN11M86 controlled by GAL10 promoter, show lethal-phenotype. But, only pAUR135 vector-removed clones as a result of low-efficient homologous recombination can grow on galactose medium preferentially.
These galactose-growing clones consist of expectedphenotype ones and reverted ones, which are AbA sensitive. So, pAUR135 is available for retransformation of these clones.
pAUR135 is effective for introduction of mutation into yeast genome and disruption of gene function. In case of transformation of industrial yeast strains, usage of pAUR135 decreases troubles with unnecessary DNA sequence.

Storage

-20°C

Concentration

0.5-1.0 µg/µl

Form

10 mM Tris-HCI (pH8.0), 1 mM EDTA

Chain length

6,074 bp

Purity

• Contains over 70% double-stranded covalently closed circular form I (RF I).
• Shown to retain cloning site by dideoxy sequencing method.
• Shown to be cleaved at a single site by restriction.

Intended usage

This is a vector for transformation of yeast S. cerevisiae using antibiotic AbA and for following removal of vector region.

Restriction enzyme map of pAUR135 DNA

Cloning sites of pAUR135 DNA