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Plasmid Files

pCMV-Tet3G

Vector encoding the Tet-On® 3G transactivator protein, which allows for inducible gene expression.

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pCMV-Tet3G Sequence and MappCMV-Tet3G.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BstZ17I (7089) AccI (7088) SfiI (6635) AvrII (6581) BmtI (6567) NheI (6563) BglII (6241) BclI * (6236) EagI (6171) MscI (5999) RsrII (5562) BstBI (5397) Bsu36I (5202) BspEI * (5071) PflMI (4923) BsgI (4690) PspXI (4209) XmnI (3816) ScaI (3697) PvuI (3587) BsrGI (47) SpeI (103) SnaBI (444) Eco53kI (670) SacI (672) SacII (759) EcoRI (766) XbaI (778) PmlI (906) PspOMI (920) ApaI (924) PpuMI (1815) BstEII (1911) HindIII (1998) Bpu10I (2021) PciI (2324) AhdI (3217) BsaI (3278) pCMV-Tet3G 7139 bp
BstZ17I  (7089)
1 site
G T A T A C C A T A T G
AccI  (7088)
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.
SfiI  (6635)
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.
AvrII  (6581)
1 site
C C T A G G G G A T C C
BmtI  (6567)
1 site
G C T A G C C G A T C G
NheI  (6563)
1 site
G C T A G C C G A T C G
BglII  (6241)
1 site
A G A T C T T C T A G A
BclI  (6236)
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.
EagI  (6171)
1 site
C G G C C G G C C G G C
MscI  (5999)
1 site
T G G C C A A C C G G T
RsrII  (5562)
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.
BstBI  (5397)
1 site
T T C G A A A A G C T T
Bsu36I  (5202)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BspEI  (5071)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PflMI  (4923)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BsgI  (4690)
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).
PspXI  (4209)
1 site
V C T C G A G B B G A G C T C V
XmnI  (3816)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3697)
1 site
A G T A C T T C A T G A
PvuI  (3587)
1 site
C G A T C G G C T A G C
BsrGI  (47)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (103)
1 site
A C T A G T T G A T C A
SnaBI  (444)
1 site
T A C G T A A T G C A T
Eco53kI  (670)
1 site
G A G C T C C T C G A G
SacI  (672)
1 site
G A G C T C C T C G A G
SacII  (759)
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.
EcoRI  (766)
1 site
G A A T T C C T T A A G
XbaI  (778)
1 site
T C T A G A A G A T C T
PmlI  (906)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PspOMI  (920)
1 site
G G G C C C C C C G G G
ApaI  (924)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PpuMI  (1815)
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.
BstEII  (1911)
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.
HindIII  (1998)
1 site
A A G C T T T T C G A A
Bpu10I  (2021)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
PciI  (2324)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (3217)
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.
BsaI  (3278)
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.
AmpR
3144 .. 4004  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3144 .. 3935  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3144 .. 4004  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3936 .. 4004  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3144 .. 4004  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
5416 .. 6210  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
5416 .. 6210  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
Tet-On® 3G
775 .. 1521  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
Tet-On® 3G
775 .. 1521  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
ori
2385 .. 2973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2385 .. 2973  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
89 .. 468  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
89 .. 468  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
6571 .. 6900  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
6571 .. 6900  =  330 bp
SV40 enhancer and early promoter
CMV promoter
469 .. 672  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
469 .. 672  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1531 .. 1665  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1531 .. 1665  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4222 .. 4356  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4222 .. 4356  =  135 bp
SV40 polyadenylation signal
AmpR promoter
4005 .. 4109  =  105 bp
AmpR promoter
4005 .. 4109  =  105 bp
small t intron
4931 .. 4996  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
4931 .. 4996  =  66 bp
simian virus 40 (SV40) small t antigen intron
SV40 ori
6585 .. 6720  =  136 bp
SV40 origin of replication
SV40 ori
6585 .. 6720  =  136 bp
SV40 origin of replication
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