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pCMV-3Tag-6

Mammalian expression vector with a hygromycin resistance marker  for tagging proteins with an N-terminal 3xFLAG epitope.

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pCMV-3Tag-6.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Agilent Technologies
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PciI (5189) BseRI (4464) AfeI (4455) PluTI (4395) SfoI (4393) NarI (4392) KasI (4391) KflI - PpuMI (4186) XcmI (4167) AsiSI (3231) BfuAI - BspMI (3180) BsmBI (2935) PshAI (2894) BstBI (2718) NsiI (5) SnaBI (346) NheI (597) BmtI (601) Eco53kI (653) SacI (655) AleI (661) BstXI (663) NotI (668) SrfI (757) BamHI (763) EcoRV (789) HindIII (793) BspDI - ClaI (800) SalI (808) AccI (809) AbsI - PaeR7I - PspXI - XhoI (814) PspOMI (823) ApaI (827) SexAI * (850) BclI * (914) BbsI (1116) SphI (1127) PsiI (1237) NgoMIV (1466) NaeI (1468) EarI (1716) XmnI (1915) DraI (1937) FspI (2292) AhdI (2515) pCMV-3Tag-6 5241 bp
PciI  (5189)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseRI  (4464)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
AfeI  (4455)
1 site
A G C G C T T C G C G A
PluTI  (4395)
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  (4393)
1 site
G G C G C C C C G C G G
NarI  (4392)
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  (4391)
1 site
G G C G C C C C G C G G
KflI  (4186)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (4186)
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.
XcmI  (4167)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
AsiSI  (3231)
1 site
G C G A T C G C C G C T A G C G
BfuAI  (3180)
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  (3180)
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.
BsmBI  (2935)
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.
PshAI  (2894)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BstBI  (2718)
1 site
T T C G A A A A G C T T
NsiI  (5)
1 site
A T G C A T T A C G T A
SnaBI  (346)
1 site
T A C G T A A T G C A T
NheI  (597)
1 site
G C T A G C C G A T C G
BmtI  (601)
1 site
G C T A G C C G A T C G
Eco53kI  (653)
1 site
G A G C T C C T C G A G
SacI  (655)
1 site
G A G C T C C T C G A G
AleI  (661)
1 site
C A C N N N N G T G G T G N N N N C A C
BstXI  (663)
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.
NotI  (668)
1 site
G C G G C C G C C G C C G G C G
SrfI  (757)
1 site
G C C C G G G C C G G G C C C G
BamHI  (763)
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.
EcoRV  (789)
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.
HindIII  (793)
1 site
A A G C T T T T C G A A
BspDI  (800)
1 site
A T C G A T T A G C T A
ClaI  (800)
1 site
A T C G A T T A G C T A
SalI  (808)
1 site
G T C G A C C A G C T G
AccI  (809)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
AbsI  (814)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (814)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (814)
1 site
V C T C G A G B B G A G C T C V
XhoI  (814)
1 site
C T C G A G G A G C T C
PspOMI  (823)
1 site
G G G C C C C C C G G G
ApaI  (827)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SexAI  (850)
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.
BclI  (914)
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.
BbsI  (1116)
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.
SphI  (1127)
1 site
G C A T G C C G T A C G
PsiI  (1237)
1 site
T T A T A A A A T A T T
NgoMIV  (1466)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (1468)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
EarI  (1716)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI recognition sequence.
Sticky ends from different EarI sites may not be compatible.
XmnI  (1915)
1 site
G A A N N N N T T C C T T N N N N A A G
DraI  (1937)
1 site
T T T A