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Plasmid Files

pCMV-3Tag-1A

Mammalian expression vector for tagging proteins with an N-terminal 3xFLAG epitope. For other reading frames, use pCMV‑3Tag-1B or pCMV‑3Tag-1C.

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pCMV-3Tag-1A.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Agilent Technologies
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PciI (4167) ApaLI (3853) BsaI (3238) PfoI (3024) BstBI (2931) RsrII (2765) BsrDI (2482) PflFI - Tth111I (2367) FspI (2351) MscI (2331) PluTI (2252) SfoI (2250) NarI (2249) KasI (2248) NdeI (240) SnaBI (346) NheI (597) BmtI (601) Eco53kI (653) SacI (655) AleI (661) SacII (662) BstXI (663) NotI (668) SrfI (757) BamHI (763) PstI (779) EcoRI (781) EcoRV (789) HindIII (793) SalI (808) AccI (809) HincII (810) AbsI - PaeR7I - PspXI - XhoI (814) PspOMI (823) ApaI (827) PvuI (905) BclI * (914) BbsI (1116) MluI (1135) PsiI (1237) DraIII (1365) SfiI (2024) BseRI (2067) StuI (2070) BsaBI * (2108) pCMV-3Tag-1A 4219 bp
PciI  (4167)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3853)
1 site
G T G C A C C A C G T G
BsaI  (3238)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
PfoI  (3024)
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.
BstBI  (2931)
1 site
T T C G A A A A G C T T
RsrII  (2765)
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.
BsrDI  (2482)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (2367)
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  (2367)
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.
FspI  (2351)
1 site
T G C G C A A C G C G T
MscI  (2331)
1 site
T G G C C A A C C G G T
PluTI  (2252)
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  (2250)
1 site
G G C G C C C C G C G G
NarI  (2249)
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  (2248)
1 site
G G C G C C C C G C G G
NdeI  (240)
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  (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
SacII  (662)
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.
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.
PstI  (779)
1 site
C T G C A G G A C G T C
EcoRI  (781)
1 site
G A A T T C C T T A A G
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
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.
HincII  (810)
1 site
G T Y R A C C A R Y T G
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.
PvuI  (905)
1 site
C G A T C G G C T A G C
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.
MluI  (1135)
1 site
A C G C G T T G C G C A
PsiI  (1237)
1 site
T T A T A