FIGURE 2: Schematic diagram of a shotgun approach to clone a small viral genome in fission yeast. (Top) the pYZ1N vector contains a wild type nmt1 promoter [68, 196]. It carries a LEU2 gene for selection. pYZ2N contains the wild type nmt1 promoter and a URA4 gene selection marker. (Bottom) pYZ3N-GFP contains the same nmt1 promoter as pYZ1N and has an added green fluorescent protein GFP tag. The α-peptide of β-galactosidase is used for selecting DNA inserts with α-complementation. Unique cloning sites in these vectors are indicated. ars1, origin of replication from S. pombe; Leu2, Saccharomyces cerevisiae leucine biosynthesis gene; AmpR, bacterial ampicillin resistance gene. Adapted from [36, 68]. Note that nothing is new in molecular features of shuttle vectors described. Goal of this figure is to illustrate a robust and streamlined strategy of shotgun gene cloning of a small viral genome.

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