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pcDNA™6.2/V5-pL-DEST

Promoterless Gateway® destination vector for cloning of mammalian promoters and genes.

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pcDNA6.2 V5-pL-DEST Sequence and MappcDNA6.2 V5-pL-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SgrDI (6577) ZraI (6576) BglI (5776) PciI (4763) BspQI - SapI (4647) PfoI (4187) BlpI (4091) NruI * (3927) BsgI (3883) PpuMI (3855) EagI (3812) BbsI (3781) BclI * (3618) AvrII (3567) StuI (3566) AatII (6578) PstI (667) BfuAI - BspMI (670) BstXI (805) BmgBI (884) SrfI (918) BbvCI (1061) BssHII (1291) BamHI (1332) EcoRI (1787) BspEI (1791) XbaI * (2414) PspOMI (2420) ApaI (2424) SacII (2427) BstBI (2431) MluI (2475) AgeI (2481) PmeI (2499) SexAI * (3334) pcDNA™6.2/V5-pL-DEST 6693 bp
SgrDI  (6577)
1 site
C G T C G A C G G C A G C T G C
ZraI  (6576)
1 site
G A C G T C C T G C A G
BglI  (5776)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
PciI  (4763)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4647)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (4647)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PfoI  (4187)
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.
BlpI  (4091)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
NruI  (3927)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
BsgI  (3883)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PpuMI  (3855)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
EagI  (3812)
1 site
C G G C C G G C C G G C
BbsI  (3781)
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.
BclI  (3618)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
AvrII  (3567)
1 site
C C T A G G G G A T C C
StuI  (3566)
1 site
A G G C C T T C C G G A
AatII  (6578)
1 site
G A C G T C C T G C A G
PstI  (667)
1 site
C T G C A G G A C G T C
BfuAI  (670)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (670)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BstXI  (805)
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.
BmgBI  (884)
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.
SrfI  (918)
1 site
G C C C G G G C C G G G C C C G
BbvCI  (1061)
1 site
C C T C A G C G G A G T C G
BssHII  (1291)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BamHI  (1332)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
EcoRI  (1787)
1 site
G A A T T C C T T A A G
BspEI  (1791)
1 site
T C C G G A A G G C C T
XbaI  (2414)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
PspOMI  (2420)
1 site
G G G C C C C C C G G G
ApaI  (2424)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SacII  (2427)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
BstBI  (2431)
1 site
T T C G A A A A G C T T
MluI  (2475)
1 site
A C G C G T T G C G C A
AgeI  (2481)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
PmeI  (2499)
1 site
G T T T A A A C C A A A T T T G
SexAI  (3334)
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
5583 .. 6443  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5583 .. 6374  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5583 .. 6443  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6375 .. 6443  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5583 .. 6443  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
1352 .. 2005  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1352 .. 2005  =  654 bp
218 amino acids  =  25.6 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
4824 .. 5412  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4824 .. 5412  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
2811 .. 3239  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2811 .. 3239  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
3696 .. 4094  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
3696 .. 4094  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
3253 .. 3582  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3253 .. 3582  =  330 bp
SV40 enhancer and early promoter
ccdB
697 .. 1002  =  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
697 .. 1002  =  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.
attR1
144 .. 268  =  125 bp
recombination site for the Gateway® LR reaction
attR1
144 .. 268  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2286 .. 2410  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2286 .. 2410  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
4252 .. 4373  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4252 .. 4373  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6444 .. 6548  =  105 bp
AmpR promoter
6444 .. 6548  =  105 bp
cat promoter
2006 .. 2108  =  103 bp
promoter of the E. coli cat gene
cat promoter
2006 .. 2108  =  103 bp
promoter of the E. coli cat gene
HSV TK poly(A) signal
2561 .. 2609  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
2561 .. 2609  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
EM7 promoter
3630 .. 3677  =  48 bp
synthetic bacterial promoter
EM7 promoter
3630 .. 3677  =  48 bp
synthetic bacterial promoter
V5 tag
2436 .. 2477  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2436 .. 2477  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
T7 promoter
96 .. 114  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
96 .. 114  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
3433 .. 3568  =  136 bp
SV40 origin of replication
SV40 ori
3433 .. 3568  =  136 bp
SV40 origin of replication
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