Fig. 4 Plasmid and primer sequences at junctions of a dual expression cassette for co-expression of eGFP and
mCherry. The eGFP and mCherry cDNAs were cloned into pKL (plasmid available from Addgene # Plasmid
#110741). PCR primers were designed to generate DNA fragments with 19- to 21-bp-long overhangs and
maintain the NheI, XhoI, BamHI, and XbaI restriction sites. The 3
0
region of the eGFP is detailed in panel (a), the
dual promoter region with the 5
0
region of the eGFP, the p10 and PH promoters, and the 5
0
region of the
mCherry cDNA in panel (b) and the 3
0
region of the mCherry cDNA in panel (c). The plasmid backbone is
amplified with the oligonucleotide pair Backbone-R/Backbone-F and the dual-promoter module with the
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Paola Rossolillo et al.