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Plasmid Files

pcDNA™6/V5-His A

Mammalian vector for expressing C-terminally V5- and 6xHis-tagged proteins. For other reading frames, use pcDNA™6/V5‑His B or pcDNA™6/V5‑His C.

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pcDNA6 V5-His A Sequence and MappcDNA6 V5-His A.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 SgrDI (5142) SspI (5025) ScaI (4701) FspI (4443) AhdI (4221) PciI (3328) BspQI - SapI (3212) BstZ17I (2949) BsmI (2897) AanI (2836) PfoI (2752) BlpI (2656) BglII (12) MfeI (161) Bpu10I (180) NdeI (484) SnaBI (590) NheI (895) BmtI (899) PmeI (904) AflII (908) HindIII (911) Acc65I (917) KpnI (921) BamHI (929) BstXI (948) EcoRI (952) PstI (961) EcoRV (964) BstXI (974) NotI (979) PaeR7I - PspXI - XhoI (985) XbaI (991) PspOMI (997) ApaI (1001) BstBI (1004) AgeI (1054) PmeI (1084) bGH poly(A) signal AanI (1734) SexAI * (1899) StuI (2131) AvrII (2132) TspMI - XmaI (2153) SmaI (2155) MscI (2264) PpuMI (2420) BsgI (2448) BsaBI * (2559) pcDNA™6/V5-His A 5144 bp
SgrDI  (5142)
1 site
C G T C G A C G G C A G C T G C
SspI  (5025)
1 site
A A T A T T T T A T A A
ScaI  (4701)
1 site
A G T A C T T C A T G A
FspI  (4443)
1 site
T G C G C A A C G C G T
AhdI  (4221)
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  (3328)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3212)
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  (3212)
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.
BstZ17I  (2949)
1 site
G T A T A C C A T A T G
BsmI  (2897)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
AanI  (2836)
2 sites
T T A T A A A A T A T T
PfoI  (2752)
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  (2656)
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.
BglII  (12)
1 site
A G A T C T T C T A G A
MfeI  (161)
1 site
C A A T T G G T T A A C
Bpu10I  (180)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
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
NheI  (895)
1 site
G C T A G C C G A T C G
BmtI  (899)
1 site
G C T A G C C G A T C G
PmeI  (904)
2 sites
G T T T A A A C C A A A T T T G
AflII  (908)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
HindIII  (911)
1 site
A A G C T T T T C G A A
Acc65I  (917)
1 site
G G T A C C C C A T G G
KpnI  (921)
1 site
G G T A C C C C A T G G
BamHI  (929)
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.
BstXI  (948)
2 sites
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.
EcoRI  (952)
1 site
G A A T T C C T T A A G
PstI  (961)
1 site
C T G C A G G A C G T C
EcoRV  (964)
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.
BstXI  (974)
2 sites
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.
NotI  (979)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (985)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (985)
1 site
V C T C G A G B B G A G C T C V
XhoI  (985)
1 site
C T C G A G G A G C T C
XbaI  (991)
1 site
T C T A G A A G A T C T
PspOMI  (997)
1 site
G G G C C C C C C G G G
ApaI  (1001)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BstBI  (1004)
1 site
T T C G A A A A G C T T
AgeI  (1054)
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  (1084)
2 sites
G T T T A A A C C A A A T T T G
AanI  (1734)
2 sites
T T A T A A A A T A T T
SexAI  (1899)
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.
StuI  (2131)
1 site
A G G C C T T C C G G A
AvrII  (2132)
1 site
C C T A G G G G A T C C
TspMI  (2153)
1 site
C C C G G G G G G C C C
XmaI  (2153)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (2155)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
MscI  (2264)
1 site
T G G C C A A C C G G T
PpuMI  (2420)
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.
BsgI  (2448)
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).
BsaBI  (2559)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
AmpR
4148 .. 5008  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4148 .. 4939  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4148 .. 5008  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4940 .. 5008  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4148 .. 5008  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3389 .. 3977  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3389 .. 3977  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1376 .. 1804  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1376 .. 1804  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
2261 .. 2659  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
2261 .. 2659  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
1818 .. 2147  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1818 .. 2147  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1106 .. 1330  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1106 .. 1330  =  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
SV40 poly(A) signal
2817 .. 2938  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2817 .. 2938  =  122 bp
SV40 polyadenylation signal
MCS
895 .. 1008  =  114 bp
multiple cloning site
MCS
895 .. 1008  =  114 bp
multiple cloning site
AmpR promoter
5009 .. 5113  =  105 bp
AmpR promoter
5009 .. 5113  =  105 bp
EM7 promoter
2195 .. 2242  =  48 bp
synthetic bacterial promoter
EM7 promoter
2195 .. 2242  =  48 bp
synthetic bacterial promoter
V5 tag
1009 .. 1050  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
1009 .. 1050  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
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
1060 .. 1077  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1060 .. 1077  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
SV40 ori
1998 .. 2133  =  136 bp
SV40 origin of replication
SV40 ori
1998 .. 2133  =  136 bp
SV40 origin of replication
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