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Plasmid Files

pCMV-3Tag-3C

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

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pCMV-3Tag-3C Sequence and MappCMV-3Tag-3C.dna
Map and Sequence File   
Sequence Author:  Agilent Technologies
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 PciI (4164) ApaLI (3850) BsaI (3235) PfoI (3021) BstBI (2928) RsrII (2762) BsrDI (2479) PflFI - Tth111I (2364) FspI (2348) MscI (2328) PluTI (2249) SfoI (2247) NarI (2246) KasI (2245) CMV enhancer 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) HincII (734) PaeR7I - PspXI - XhoI (740) PspOMI (820) ApaI (824) stop codons PvuI (902) BclI * (911) bGH poly(A) signal BbsI (1113) MluI (1132) PsiI (1234) DraIII (1362) SfiI (2021) BseRI (2064) StuI (2067) BsaBI * (2105) pCMV-3Tag-3C 4216 bp
PciI  (4164)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3850)
1 site
G T G C A C C A C G T G
BsaI  (3235)
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  (3021)
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  (2928)
1 site
T T C G A A A A G C T T
RsrII  (2762)
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  (2479)
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  (2364)
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  (2364)
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  (2348)
1 site
T G C G C A A C G C G T
MscI  (2328)
1 site
T G G C C A A C C G G T
PluTI  (2249)
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  (2247)
1 site
G G C G C C C C G C G G
NarI  (2246)
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  (2245)
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  (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.
HincII  (734)
1 site
G T Y R A C C A R Y T G
PaeR7I  (740)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (740)
1 site
V C T C G A G B B G A G C T C V
XhoI  (740)
1 site
C T C G A G G A G C T C
PspOMI  (820)
1 site
G G G C C C C C C G G G
ApaI  (824)
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  (902)
1 site
C G A T C G G C T A G C
BclI  (911)
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  (1113)
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  (1132)
1 site
A C G C G T T G C G C A
PsiI  (1234)
1 site
T T A T A A A A T A T T
DraIII  (1362)
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  (2021)
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  (2064)
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.
StuI  (2067)
1 site
A G G C C T T C C G G A
BsaBI  (2105)
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.
NeoR/KanR
2118 .. 2912  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2118 .. 2912  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
3520 .. 4108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3520 .. 4108  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1138 .. 1593  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1138 .. 1593  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1726 .. 2083  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1726 .. 2083  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
66 .. 370  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
66 .. 370  =  305 bp
human cytomegalovirus immediate early enhancer
bGH poly(A) signal
924 .. 1131  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
924 .. 1131  =  208 bp
bovine growth hormone polyadenylation signal
CMV promoter
371 .. 574  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
371 .. 574  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
AmpR promoter
1620 .. 1722  =  103 bp
AmpR promoter
1620 .. 1722  =  103 bp
MCS
651 .. 745  =  95 bp
multiple cloning site
MCS
651 .. 745  =  95 bp
multiple cloning site
3xFLAG
746 .. 817  =  72 bp
24 amino acids  =  3.0 kDa
Product: three tandem FLAG epitope tags
3xFLAG
746 .. 817  =  72 bp
24 amino acids  =  3.0 kDa
Product: three tandem FLAG epitope tags
HSV TK poly(A) signal
3144 .. 3191  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3144 .. 3191  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
T3 promoter
620 .. 638  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
620 .. 638  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T7 promoter
871 .. 889  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
871 .. 889  =  19 bp
promoter for bacteriophage T7 RNA polymerase
stop codons
833 .. 843  =  11 bp
stop codons in all three reading frames
stop codons
833 .. 843  =  11 bp
stop codons in all three reading frames
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