EUCOMM and KOMP-CSD Gene Targeting Cassettes

Promoter-Containing Targeting Cassettes

L1L2_Bact_P The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter Reading frame-independent LacZ gene trap cassette: En2 splice acceptor / ECMV IRES / LacZ / SV40 polyadenylation site — Selectable Marker Human β-actin promoter driving the wild-type neoR gene: human β-actin promoter / neomycin phosphotransferase (neoR) gene / SV40 polyA — Plasmid High copy number, ClonNAT resistant (25ug/ml). No restrictions on E. Coli host strain. Also weakly Kan resistant(20ug/ml), presumably from a cryptic bacterial promoter. —

L1L2_PGK_P The targeting cassette is flanked by Gateway L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter Reading frame-independent LacZ gene trap cassette: En2 splice acceptor / ECMV IRES /LacZ / SV40 polyadenylation site — Selectable Marker Phosphoglycerate Kinase 1 (PGK1) promoter driving the wild-type neoR gene: PGK promoter / neomycin phosphotransferase (neoR) gene / SV40 polyA — Plasmid High copy number, ClonNAT resistant (25ug/ml). Kan resistant(100ug/ml) from bacterial EM7 promoter upstream of neo. No restrictions on E. Coli host strain. —

Promoterless Targeting Cassettes

L1L2_gt0 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless, (reading frame 0) gene trap cassette. LacZ reporter gene and WT neomycin phosphotranferase (neoR) gene are translated from T2A oligopeptides1 as discreet proteins, not as fusions: En-2 splice acceptor (frame 0) / T2A / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant (25ug/ml). No restrictions on E. Coli host strain. —

L1L2_gt1 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless, (reading frame 1) gene trap cassette. LacZ reporter gene and WT neomycin phosphotranferase (neoR) gene are translated from T2A oligopeptides1 as discreet proteins, not as fusions: En-2 splice acceptor (frame 1) / T2A / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant (25ug/ml). No restrictions on E. Coli host strain. —

L1L2_gt2 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless, (reading frame 2) gene trap cassette. LacZ reporter gene and WT neomycin phosphotranferase (neoR) gene are translated from T2A oligopeptides1 as discreet proteins, not as fusions: En-2 splice acceptor (frame 2) / T2A / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant (25ug/ml). No restrictions on E. Coli host strain. —

L1L2_gtK The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless gene trap cassette containing an initiating ATG and optimal Kozak translation sequence. To be used as first coding exon downstream of untranslated exons lacking an initiating ATG. LacZ reporter gene and WT neomycin phosphotranferase (neoR) gene are translated from T2A oligopeptides1 as discreet proteins, not as fusions: En-2 splice acceptor (Kozak/ATG)/ T2A / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant (25ug/ml). No restrictions on E. Coli host strain. —

L1L2_st0 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless (reading frame 0) secretory trap cassette2 containing rat CD4 transmembrane (Tm) domain fused to N terminus of a lacZ reporter. WT neomycin phosphotranferase (neoR) gene is translated from a T2A oligopeptide1 as a discreet protein, not as a fusion protein: En-2 splice acceptor (frame 0) / ratCD4 Tm domain / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant(25ug/ml). No restrictions on E.Coli host strain. —

L1L2_st1 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless (reading frame 1) secretory trap cassette2 containing rat CD4 transmembrane (Tm) domain fused to N terminus of a lacZ reporter. WT neomycin phosphotranferase (neoR) gene is translated from a T2A oligopeptide1 as a discreet protein, not as a fusion protein: En-2 splice acceptor (frame 1) / ratCD4 Tm domain / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant(25ug/ml). No restrictions on E.Coli host strain. —

L1L2_st2 The targeting cassette is flanked by L1/L2 Gateway sites for insertion into R1/R2 Gateway sites upstream of the critical exon(s) in the intermediate vector.

Reporter / Selectable Marker Promoterless (reading frame 2) secretory trap cassette2 containing rat CD4 transmembrane (Tm) domain fused to N terminus of a lacZ reporter. WT neomycin phosphotranferase (neoR) gene is translated from a T2A oligopeptide1 as a discreet protein, not as a fusion protein: En-2 splice acceptor (frame 2) / ratCD4 Tm domain / LacZ / T2A / neoR / SV40 polyadenylation site — Plasmid High copy number, ClonNAT resistant(25ug/ml). No restrictions on E.Coli host strain. —