Asp718 I TOPO

EcoR V
BamH I
Hind III

BstX I

Pme I
Sac II
Kpn I

Xho I
Xba I
Apa I

Age I
Not I
T7 CCC TT AAG GGT
GGG AAG TGG TTC CCA V5 epitope 6xHis Stop

TOPO
BGH pA
V
P CM

SV
40 o
pcDNA™3.1D/

ri
A m p i c i l li

Neomy
V5-His-TOPO®
5514 bp

cin
n

SV
40
Comments for pcDNA™3.1D/V5-His-TOPO® A p
5514 nucleotides p U C o ri
CMV promoter: bases 232-819
T7 promoter/priming site: bases 863-882
TOPO® recognition site 1: bases 930-934
Overhang sequence (complementary strand): bases 935-938
TOPO® recognition site 2: bases 939-943
V5 epitope: bases 1011-1052
Polyhistidine (6xHis) tag: bases 1062-1079
BGH reverse priming site: bases 1102-1119
BGH polyadenylation signal: bases 1108-1332
SV40 early promoter and origin: bases 1833-2142
Neomycin resistance gene: bases 2217-3011
SV40 early polyadenylation signal: bases 3189-3319
pUC origin: bases 3700-4373 (complementary strand)
Ampicillin (bla) resistance gene: bases 4518-5378 (complementary strand)
bla promoter: bases 5379-5477 (complementary strand)