The pYSG-IBAwt1 vector is designed for high-level expression of target proteins without affinity tag in yeast.
The vector carries an ampicillin resistance cassette for selection of transformed E. coli cells, the ColE1 origin for a high plasmid copy number, the copper-inducible promoter (CUP1) for controlled high-level expression, the URA3 auxotrophy marker for selection after transformation (do not use URA3 for selection during expression), the LEU2d auxotrophy marker for selection to increase plasmid copy number for expression (do not use LEU2d for selection after transformation), and the 2 micron ori for episomal replication in yeast.
Please note that cloning into
pYSG-IBA Acceptor Vectors compulsorily requires the restriction enzyme
Esp3I since no other MCS for the integration of a gene of interest is available. In addition to the
direct cloning of the gene of interest into pYSG-IBA vectors with Esp3I, another option via a so-called
Entry Vector is possible.