pcDNA6 V5-His C

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

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (5138) SspI (5021) ScaI (4697) FspI (4439) AhdI (4217) PciI (3324) BspQI - SapI (3208) BstZ17I (2945) BsmI (2893) PfoI (2748) BlpI (2652) BglII (12) MfeI (161) Bpu10I (180) NdeI (484) SnaBI (590) NheI (895) BmtI (899) AflII (908) HindIII (911) Acc65I (917) KpnI (921) BamHI (929) EcoRI (952) PstI (961) EcoRV (964) NotI (979) PaeR7I - PspXI - XhoI (985) BstEII (990) BstBI (1000) AgeI (1050) 6xHis CsiI - SexAI * (1895) StuI (2127) AvrII (2128) TspMI - XmaI (2149) SmaI (2151) MscI (2260) EcoO109I - PpuMI (2416) BsgI (2444) BsaBI * (2555) pcDNA™6/V5-His C 5140 bp
SgrDI  (5138)
1 site
C G T C G A C G G C A G C T G C
SspI  (5021)
1 site
A A T A T T T T A T A A
ScaI  (4697)
1 site
A G T A C T T C A T G A
FspI  (4439)
1 site
T G C G C A A C G C G T
AhdI  (4217)
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  (3324)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3208)
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  (3208)
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  (2945)
1 site
G T A T A C C A T A T G
BsmI  (2893)
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.
PfoI  (2748)
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  (2652)
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

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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
AflII  (908)
1 site
C T T A A G G A A T T C
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.
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.
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
BstEII  (990)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BstBI  (1000)
1 site
T T C G A A A A G C T T
AgeI  (1050)
1 site
A C C G G T T G G C C A
CsiI  (1895)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (1895)
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  (2127)
1 site
A G G C C T T C C G G A
AvrII  (2128)
1 site
C C T A G G G G A T C C
TspMI  (2149)
1 site
C C C G G G G G G C C C
XmaI  (2149)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2151)
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  (2260)
1 site
T G G C C A A C C G G T
EcoO109I  (2416)
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.
PpuMI  (2416)
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  (2444)
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  (2555)
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
4144 .. 5004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4144 .. 4935  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4144 .. 5004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4936 .. 5004  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4144 .. 5004  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3385 .. 3973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3385 .. 3973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1372 .. 1800  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1372 .. 1800  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
2257 .. 2655  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
2257 .. 2655  =  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
1814 .. 2143  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1814 .. 2143  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1102 .. 1326  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1102 .. 1326  =  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
2813 .. 2934  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2813 .. 2934  =  122 bp
SV40 polyadenylation signal
MCS
895 .. 1004  =  110 bp
multiple cloning site
MCS
895 .. 1004  =  110 bp
multiple cloning site
AmpR promoter
5005 .. 5109 &