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The pYSG-IBA164 vector is designed for high-level expression of target proteins with an N-terminal Twin-Strep-tag® and C-terminal FLAG-tag in yeast. The vector carries an ampicillin resistance cassette for selection of transformed E. coli cells, the copper-inducible promoter (CUP1) for controlled high-level expression, the LEU2d auxotrophy marker for selection to increase plasmid copy number for expression (do not use LEU2d for selection after transformation), the ColE1 origin for a high plasmid copy number, the URA3 auxotrophy marker is for the selection after transformation (do not use URA3 for selection during expression), 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 directly cloning the gene of interest into pYSG-IBA vectors with Esp3I, another option via an Entry Vector is possible.
