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pCMV-Tet3G

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

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pCMV-Tet3G.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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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
ORF:  628 .. 1521  =  894 bp
ORF:  297 amino acids  =  33.8 kDa
ORF:  1756 .. 2157  =  402 bp
ORF:  133 amino acids  =  14.7 kDa
ORF:  3274 .. 3540  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5365 .. 5901  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  932 .. 1234  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  1934 .. 2284  =  351 bp
ORF:  116 amino acids  =  13.3 kDa
ORF:  5120 .. 5365  =  246 bp
ORF:  81 amino acids  =  8.4 kDa
ORF:  1227 .. 1682  =  456 bp
ORF:  151 amino acids  =  16.0 kDa
ORF:  3144 .. 4004  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  5652 .. 6038  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  4379 .. 4852  =  474 bp
ORF:  157 amino acids  =  18.2 kDa
ORF:  5033 .. 5395  =  363 bp
ORF:  120 amino acids  =  13.6 kDa
ORF:  1630 .. 1962  =  333 bp
ORF:  110 amino acids  =  12.0 kDa
ORF:  5416 .. 6210  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
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