pCMV-Tag 5B

Mammalian expression vector for tagging proteins with a C-terminal Myc epitope. For other reading frames, use pCMV-Tag 5A or pCMV-Tag 5C.

Sequence Author: Agilent Technologies

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PciI (4271) ApaLI (3957) BsaI (3342) PfoI (3128) BstBI (3035) RsrII (2869) BsrDI (2586) PflFI - Tth111I (2471) MscI (2435) PluTI (2356) SfoI (2354) NarI (2353) KasI (2352) StuI (2174) NdeI (240) SnaBI (346) NheI (597) BmtI (601) Eco53kI (653) SacI (655) AleI (661) SacII (662) BstXI (663) NotI (668) SrfI (682) BamHI (687) PstI (703) EcoRI (705) EcoRV (713) HindIII (717) SalI (732) AccI (733) PaeR7I - PspXI - XhoI (739) EarI (755) PspOMI (774) ApaI (778) PvuI (856) BclI * (1010) MfeI (1103) HpaI (1116) BtsI - BtsαI (1192) MluI (1239) DraIII (1469) SfiI (2128) BseRI (2171) pCMV-Tag 5B 4323 bp
PciI  (4271)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3957)
1 site
G T G C A C C A C G T G
BsaI  (3342)
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  (3128)
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  (3035)
1 site
T T C G A A A A G C T T
RsrII  (2869)
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  (2586)
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  (2471)
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  (2471)
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.
MscI  (2435)
1 site
T G G C C A A C C G G T
PluTI  (2356)
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  (2354)
1 site
G G C G C C C C G C G G
NarI  (2353)
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  (2352)
1 site
G G C G C C C C G C G G
StuI  (2174)
1 site
A G G C C T T C C G G A
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  (682)
1 site
G C C C G G G C C G G G C C C G
BamHI  (687)
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  (703)
1 site
C T G C A G G A C G T C
EcoRI  (705)
1 site
G A A T T C C T T A A G
EcoRV  (713)
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  (717)
1 site
A A G C T T T T C G A A
SalI  (732)
1 site
G T C G A C C A G C T G
AccI  (733)
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.
PaeR7I  (739)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (739)
1 site
V C T C G A G B B G A G C T C V
XhoI  (739)
1 site
C T C G A G G A G C T C
EarI  (755)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
PspOMI  (774)
1 site
G G G C C C C C C G G G
ApaI  (778)
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  (856)
1 site
C G A T C G G C T A G C
BclI  (1010)
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.
MfeI  (1103)
1 site
C A A T T G G T T A A C
HpaI  (1116)
1 site
G T T A A C C A A T T G
BtsI  (1192)
1 site
G C A G T G N N C G T C A C
BtsαI  (1192)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
MluI  (1239)
1 site
A C G C G T T G C G C A
DraIII  (1469)
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.
SfiI  (2128)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
BseRI  (2171)
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.
NeoR/KanR
2225 .. 3019  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2225 .. 3019  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
3627 .. 4215  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3627 .. 4215  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1245 .. 1700  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1245 .. 1700  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1833 .. 2190  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1833 .. 2190  =  358 bp
SV40 enhancer and early promoter
CMV en