The vector pKLMF-EK directs high-level intracellular expression of a recombinant protein fused to the maltose binding protein (MBP) in the yeast Kluyveromyces lactis. It is compatible with the K. lactis Protein Expression Kit (NEB #E1000). MBP fusion proteins expressed from pKLMF-EK can be affinity purified from cell lysates using Amylose Resin (NEB #E8021). MBP can be removed from purified fusion proteins by digestion with Enterokinase (NEB #P8070).
Vector pKLMF-EK contains the strong K. lactis PLAC4-PBI promoter (1), DNA encoding E. coli maltose binding protein (MBP) with a C-terminal enterokinase site, a universal multiple cloning site (MCS), the K. lactis LAC4 transcription terminator (TT), and a fungal acetamidase selectable marker gene (amdS) expressed from the yeast ADH1 promoter (PADH1). An E. coli replication origin (ori) and ampicillin resistance gene (ApR) is present for propagation of pKLMF-EK in E. coli. SacII linearized pKLMF-EK integrates into the LAC4 locus of the K. lactis genome upon transformation of K. lactis competent cells.
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