pCMV6-AC-GFP-rtTA

Vector for efficient doxycycline-inducible expression of a C-terminally TurboGFP-tagged protein.

Sequence Author: OriGene

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DraIII (8570) AlwNI (6944) PciI (6528) SalI (6154) BstZ17I (6149) BsmI (6097) PfoI (5952) BstBI (5859) PflFI - Tth111I (5295) BclI * (5018) AvrII (4967) SfiI (4920) CsiI - SexAI * (4734) XcmI (4642) AgeI (4547) BsrGI (301) SpeI (357) CMV enhancer SnaBI (698) EcoRI (979) XbaI (988) NruI (1265) MfeI (1502) BamHI (1732) HpaI (2154) BsiWI (2337) BspEI * (2537) FspAI (2749) BglII (2776) Acc65I (3167) KpnI (3171) AsiSI (3193) MreI - SgrAI (3195) AscI (3199) HindIII (3213) NheI (3224) BmtI (3228) RsrII (3230) MluI (3236) NotI (3245) PaeR7I - XhoI (3251) PaqCI (3708) PmeI (3959) SacII (3973) AleI (4059) BbvCI (4285) BsmBI - Esp3I (4349) pCMV6-AC-GFP-rtTA 8767 bp
DraIII  (8570)
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.
AlwNI  (6944)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PciI  (6528)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (6154)
1 site
G T C G A C C A G C T G
BstZ17I  (6149)
1 site
G T A T A C C A T A T G
BsmI  (6097)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
PfoI  (5952)
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  (5859)
1 site
T T C G A A A A G C T T
PflFI  (5295)
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  (5295)
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.
BclI  (5018)
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.
AvrII  (4967)
1 site
C C T A G G G G A T C C
SfiI  (4920)
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.
CsiI  (4734)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (4734)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
XcmI  (4642)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
AgeI  (4547)
1 site
A C C G G T T G G C C A
BsrGI  (301)
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  (357)
1 site
A C T A G T T G A T C A
SnaBI  (698)
1 site
T A C G T A A T G C A T
EcoRI  (979)
1 site
G A A T T C C T T A A G
XbaI  (988)
1 site
T C T A G A A G A T C T
NruI  (1265)
1 site
T C G C G A A G C G C T
MfeI  (1502)
1 site
C A A T T G G T T A A C
BamHI  (1732)
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.
HpaI  (2154)
1 site
G T T A A C C A A T T G
BsiWI  (2337)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BspEI  (2537)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
FspAI  (2749)
1 site
R T G C G C A Y Y A C G C G T R
BglII  (2776)
1 site
A G A T C T T C T A G A
Acc65I  (3167)
1 site
G G T A C C C C A T G G
KpnI  (3171)
1 site
G G T A C C C C A T G G
AsiSI  (3193)
1 site
G C G A T C G C C G C T A G C G
MreI  (3195)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3195)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
AscI  (3199)
1 site
G G C G C G C C C C G C G C G G
HindIII  (3213)
1 site
A A G C T T T T C G A A
NheI  (3224)
1 site
G C T A G C C G A T C G
BmtI  (3228)
1 site
G C T A G C C G A T C G
RsrII  (3230)
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.
MluI  (3236)
1 site
A C G C G T T G C G C A
NotI  (3245)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (3251)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3251)
1 site
C T C G A G G A G C T C
PaqCI  (3708)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
PmeI  (3959)
1 site
G T T T A A A C C A A A T T T G
SacII  (3973)
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.
AleI  (4059)
1 site
C A C N N N N G T G G T G N N N N C A C
BbvCI  (4285)
1 site
C C T C A G C G G A G T C G
BsmBI  (4349)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (4349)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
AmpR
7345 .. 8205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7345 .. 8136  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7345 .. 8205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8137 .. 8205  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7345 .. 8205  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
5049 .. 5843  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
5049 .. 5843  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 1:  rTetR  
   985 .. 1602  =  618 bp
   206 amino acids  =  23.2 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 2:  
   1603 .. 1608  =  6 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 3:  VP16 minimal transcriptional activation domain  
   1609 .. 1644  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 4:  
   1645 .. 1647  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 5:  VP16 minimal transcriptional activation domain  
   1648 .. 1683  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 6:  
   1684 .. 1686  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 7:  VP16 minimal transcriptional activation domain  
   1687 .. 1722  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 8:  
   1723 .. 1731  =  9 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
985 .. 1731  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
TurboGFP
3257 .. 3952  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
3257 .. 3952  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
hGH poly(A) signal
4002 .. 4624  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
4002 .. 4624  =  623 bp
human growth hormone polyadenylation signal
ori
6589 .. 7174  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6589 .. 7174  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
EF-1α poly(A) signal
1762 .. 2334  =  573 bp
human EF-1α polyadenylation signal
EF-1α poly(A) signal
1762 .. 2334  =  573 bp
human EF-1α polyadenylation signal
f1 ori
8337 .. 8765  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
8337 .. 8765  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
343 .. 722  =  380 bp
human cytomegalovirus immediate early enhancer
TRE3G promoter
2782 .. 3160  =  379 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G
TRE3G promoter
2782 .. 3160  =  379 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G
SV40 promoter
4653 .. 4982  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4653 .. 4982  =  330 bp
SV40 enhancer and early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
723 .. 926  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
6017 .. 6138  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6017 .. 6138  =  122 bp
SV40 polyadenylation signal
AmpR promoter
8206 .. 8310  =  105 bp
AmpR promoter
8206 .. 8310  =  105 bp
MCS
3167 .. 3256  =  90 bp
multiple cloning site
MCS
3167 .. 3256  =  90 bp
multiple cloning site
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
952 .. 970  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4833 .. 4968  =  136 bp
SV40 origin of replication
SV40 ori
4833 .. 4968  =  136 bp
SV40 origin of replication
tet operator
2790 .. 2808  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2790 .. 2808  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2826 .. 2844  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2826 .. 2844  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2862 .. 2880  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2862 .. 2880  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2898 .. 2916  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2898 .. 2916  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2934 .. 2952  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2934 .. 2952  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2970 .. 2988  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2970 .. 2988  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3006 .. 3024  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3006 .. 3024  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  985 .. 1731  =  747 bp
ORF:  248 amino acids  =  27.8 kDa
ORF:  2356 .. 2691  =  336 bp
ORF:  111 amino acids  =  12.8 kDa
ORF:  5221 .. 5607  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1142 .. 1444  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  1652 .. 1957  =  306 bp
ORF:  101 amino acids  =  11.4 kDa
ORF:  2813 .. 3079  =  267 bp
ORF:  88 amino acids  =  10.4 kDa
ORF:  3257 .. 3961  =  705 bp
ORF:  234 amino acids  =  25.9 kDa
ORF:  7475 .. 7741  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3729 .. 3992  =  264 bp
ORF:  87 amino acids  =  9.1 kDa
ORF:  5049 .. 5843  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3221 .. 4102  =  882 bp
ORF:  293 amino acids  =  29.1 kDa
ORF:  2014 .. 2388  =  375 bp
ORF:  124 amino acids  =  14.1 kDa
ORF:  4366 .. 4590  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  7345 .. 8205  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1437 .. 1718  =  282 bp
ORF:  93 amino acids  =  9.5 kDa
ORF:  5358 .. 5894  =  537 bp
ORF:  178 amino acids  =  19.7 kDa
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Download pCMV6-AC-GFP-rtTA.dna file

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Individual Sequences & Maps

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