Skip to main content
. Author manuscript; available in PMC: 2018 Jun 1.
Published in final edited form as: Curr Genet. 2016 Oct 25;63(3):519–530. doi: 10.1007/s00294-016-0659-2

Fig. 1.

Fig. 1

Construction of two plasmids for transformation of multiple microbes. A. Plasmid or DNA maps and the construction process. In step 1, the S. cerevisiae 2 micron circle and URA3 gene were amplified from pRS426 with primers ALID2018–ALID2019 and cloned into the ScaI site of pPZP-201BK, to form plasmid pGI3. In step 2, the NAT cassette was amplified from pPZP-NATcc with primers ai290–ALID2147 and the GAL7 promoter from C. neoformans genomic DNA with primers GI114-GI115, and cloned into pGI3 to form pGI7. AmpR and KanR refer to genes conferring resistance to ampicillin and kanamycin in bacteria. For simplicity, replication components in E. coli and A. tumefaciens are omitted. Tick marks indicate 100 bp intervals. B. Sequence of the GI114-GI115 amplicon, including the 1,020 bp immediately upstream of the GAL7 open reading frame. Sequences corresponding to primers GI114 and GI115 are underlined. The right border sequence in the T-DNA is in bold, with the three nucleotides that usually integrate into the fungal genome in the box with the grey highlight.