pCMV-DDK-N

Mammalian vector for CMV promoter-driven expression of an ORF with an N-terminal FLAG® tag.

Sequence Author: transOMIC

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HindIII (853) MscI (862) SfiI (868) EcoRI (875) SalI (882) AccI (883) BglII (890) AvaI - BmeT110I - BsoBI - PaeR7I - XhoI (896) Acc65I (901) KpnI (905) EagI - NotI (908) AanI (1032) M13 rev pCMV-DDK-N 3779 bp
HindIII  (853)
1 site
A A G C T T T T C G A A
MscI  (862)
1 site
T G G C C A A C C G G T
SfiI  (868)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
EcoRI  (875)
1 site
G A A T T C C T T A A G
SalI  (882)
1 site
G T C G A C C A G C T G
AccI  (883)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BglII  (890)
1 site
A G A T C T T C T A G A
AvaI  (896)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BmeT110I  (896)
1 site
C Y C G R G G R G C Y C
BsoBI  (896)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (896)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (896)
1 site
C T C G A G G A G C T C
Acc65I  (901)
1 site
G G T A C C C C A T G G
KpnI  (905)
1 site
G G T A C C C C A T G G
EagI  (908)
1 site
C G G C C G G C C G G C
NotI  (908)
1 site
G C G G C C G C C G C C G G C G
AanI  (1032)
1 site
T T A T A A A A T A T T
AmpR
2324 .. 3184  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2324 .. 3115  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2324 .. 3184  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3116 .. 3184  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2324 .. 3184  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1565 .. 2153  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
1565 .. 2153  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 330  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
331 .. 534  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
918 .. 1052  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
918 .. 1052  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3185 .. 3289  =  105 bp
AmpR promoter
3185 .. 3289  =  105 bp
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
SV40 intron
672 .. 768  =  97 bp
modified SV40 intron with splice donor and acceptor sites
MCS
853 .. 914  =  62 bp
multiple cloning site
MCS
853 .. 914  =  62 bp
multiple cloning site
M13 rev
1163 .. 1179  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
1163 .. 1179  =  17 bp
common sequencing primer, one of multiple similar variants
ATG
829 .. 831  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
829 .. 831  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
FLAG
832 .. 855  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
832 .. 855  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
ORF:  2454 .. 2720  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3570 .. 36  =  246 bp
ORF:  81 amino acids  =  9.5 kDa
ORF:  3530 .. 113  =  363 bp
ORF:  120 amino acids  =  13.4 kDa
ORF:  2324 .. 3184  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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