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pcDNA™3.1/nV5-DEST

Gateway® destination vector for expressing N-terminally V5-tagged proteins in mammalian cells.

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pcDNA3.1 nV5-DEST Sequence and MappcDNA3.1 nV5-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 BglII (12) PvuI (6583) AhdI (6213) PciI (5323) PfoI (4747) BstBI (4654) RsrII (4488) PflFI - Tth111I (4090) PluTI (3975) SfoI (3973) NarI (3972) KasI (3971) AvrII (3762) StuI (3761) MfeI (161) NruI (208) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) HindIII (902) ATG TEV Site EcoO109I - PspOMI (973) ApaI (977) EcoRV (1003) NotI (1133) lac UV5 promoter BspEI (1449) BsmBI (1677) PasI - PflMI * (1685) BbvCI (2178) SrfI (2324) BmgBI (2358) BstXI (2441) BbsI (2925) DraIII (3239) SexAI * (3529) pcDNA™3.1/nV5-DEST 7136 bp
BglII  (12)
1 site
A G A T C T T C T A G A
PvuI  (6583)
1 site
C G A T C G G C T A G C
AhdI  (6213)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PciI  (5323)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PfoI  (4747)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (4654)
1 site
T T C G A A A A G C T T
RsrII  (4488)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
PflFI  (4090)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4090)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PluTI  (3975)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (3973)
1 site
G G C G C C C C G C G G
NarI  (3972)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
KasI  (3971)
1 site
G G C G C C C C G C G G
AvrII  (3762)
1 site
C C T A G G G G A T C C
StuI  (3761)
1 site
A G G C C T T C C G G A
MfeI  (161)
1 site
C A A T T G G T T A A C
NruI  (208)
1 site
T C G C G A A G C G C T
SpeI  (249)
1 site
A C T A G T T G A T C A
NdeI  (484)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SnaBI  (590)
1 site
T A C G T A A T G C A T
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A G
HindIII  (902)
1 site
A A G C T T T T C G A A
EcoO109I  (973)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (973)
1 site
G G G C C C C C C G G G
ApaI  (977)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoRV  (1003)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
NotI  (1133)
1 site
G C G G C C G C C G C C G G C G
BspEI  (1449)
1 site
T C C G G A A G G C C T
BsmBI  (1677)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PasI  (1685)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (1685)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BbvCI  (2178)
1 site
C C T C A G C G G A G T C G
SrfI  (2324)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2358)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BstXI  (2441)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BbsI  (2925)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
DraIII  (3239)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
SexAI  (3529)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AmpR
6140 .. 7000  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6140 .. 6931  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6140 .. 7000  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6932 .. 7000  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6140 .. 7000  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
3844 .. 4638  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
3844 .. 4638  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
CmR
1240 .. 1899  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1240 .. 1899  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5384 .. 5969  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5384 .. 5969  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3006 .. 3434  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3006 .. 3434  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
3448 .. 3777  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3448 .. 3777  =  330 bp
SV40 enhancer and early promoter
ccdB
2241 .. 2546  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
ccdB
2241 .. 2546  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA
gyrase
Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains.
bGH poly(A) signal
2736 .. 2960  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
2736 .. 2960  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
attR1
1007 .. 1131  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1007 .. 1131  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2587 .. 2711  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2587 .. 2711  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
4812 .. 4933  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4812 .. 4933  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7001 .. 7105  =  105 bp
AmpR promoter
7001 .. 7105  =  105 bp
V5 tag
914 .. 955  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
914 .. 955  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
1156 .. 1186  =  31 bp
   Segment 1:  -35  
   1156 .. 1161  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1156 .. 1186  =  31 bp
   Segment 2:  
   1162 .. 1179  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1156 .. 1186  =  31 bp
   Segment 3:  -10  
   1180 .. 1186  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1156 .. 1186  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
TEV Site
956 .. 976  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) recognition and
cleavage site
TEV Site
956 .. 976  =  21 bp
7 amino acids  =  870.0 Da
Product: tobacco etch virus (TEV) recognition and
cleavage site
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ATG
911 .. 913  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
911 .. 913  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
3628 .. 3763  =  136 bp
SV40 origin of replication
SV40 ori
3628 .. 3763  =  136 bp
SV40 origin of replication
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