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Plasmid Files

pcDNA™-DEST40

Gateway® destination vector for expressing C-terminally V5- and 6xHis-tagged proteins in mammalian cells.

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pcDNA-DEST40 Sequence and MappcDNA-DEST40.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SgrDI (7141) PvuI (6590) AhdI (6220) PciI (5327) PfoI (4751) RsrII (4490) PflFI - Tth111I (4092) PluTI (3977) SfoI (3975) NarI (3974) KasI (3973) AvrII (3764) StuI (3763) MfeI (161) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) NotI (1037) BspEI (1353) EcoRI (1357) BsmBI (1581) PasI - PflMI * (1589) BbvCI (2082) SrfI (2228) BmgBI (2262) BstXI (2345) XbaI * (2619) EcoO109I - PspOMI (2625) ApaI (2629) SacII (2632) BstBI (2636) AgeI (2686) PmeI (2716) bGH poly(A) signal BbsI (2927) DraIII (3241) SexAI * (3531) pcDNA™-DEST40 7143 bp
SgrDI  (7141)
1 site
C G T C G A C G G C A G C T G C
PvuI  (6590)
1 site
C G A T C G G C T A G C
AhdI  (6220)
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  (5327)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PfoI  (4751)
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.
RsrII  (4490)
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  (4092)
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  (4092)
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  (3977)
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  (3975)
1 site
G G C G C C C C G C G G
NarI  (3974)
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  (3973)
1 site
G G C G C C C C G C G G
AvrII  (3764)
1 site
C C T A G G G G A T C C
StuI  (3763)
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
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
NotI  (1037)
1 site
G C G G C C G C C G C C G G C G
BspEI  (1353)
1 site
T C C G G A A G G C C T
EcoRI  (1357)
1 site
G A A T T C C T T A A G
BsmBI  (1581)
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  (1589)
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  (1589)
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  (2082)
1 site
C C T C A G C G G A G T C G
SrfI  (2228)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (2262)
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  (2345)
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.
XbaI  (2619)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
EcoO109I  (2625)
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  (2625)
1 site
G G G C C C C C C G G G
ApaI  (2629)
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  (2632)
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  (2636)
1 site
T T C G A A A A G C T T
AgeI  (2686)
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  (2716)
1 site
G T T T A A A C C A A A T T T G
BbsI  (2927)
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  (3241)
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  (3531)
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
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6147 .. 6938  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6939 .. 7007  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6147 .. 7007  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
3846 .. 4640  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
3846 .. 4640  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
CmR
1144 .. 1803  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1144 .. 1803  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
5388 .. 5976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5388 .. 5976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3008 .. 3436  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3008 .. 3436  =  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
3450 .. 3779  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3450 .. 3779  =  330 bp
SV40 enhancer and early promoter
ccdB
2145 .. 2450  =  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
2145 .. 2450  =  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
2738 .. 2962  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
2738 .. 2962  =  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
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR1
911 .. 1035  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2491 .. 2615  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2491 .. 2615  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
4816 .. 4937  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4816 .. 4937  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7008 .. 7112  =  105 bp
AmpR promoter
7008 .. 7112  =  105 bp
V5 tag
2641 .. 2682  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2641 .. 2682  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
1060 .. 1090  =  31 bp
   Segment 1:  -35  
   1060 .. 1065  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
   Segment 2:  
   1066 .. 1083  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
   Segment 3:  -10  
   1084 .. 1090  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
1060 .. 1090  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
2692 .. 2709  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2692 .. 2709  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
SV40 ori
3630 .. 3765  =  136 bp
SV40 origin of replication
SV40 ori
3630 .. 3765  =  136 bp
SV40 origin of replication
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