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Plasmid Files

pRetroX-TetOne™

All-in-one retroviral vector for strong inducible expression of genes using the Tet-On® system.

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pRetroX-TetOne Sequence and MappRetroX-TetOne.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BspQI - SapI (8267) PciI (8150) AlwNI (7741) ScaI (6781) SspI (6457) SacII (5120) AleI (5119) MluI (5092) EcoRV (4734) PflMI (4557) BsiWI (4433) AvrII (77) SnaBI (519) AfeI (717) Eco53kI (733) SacI (735) PshAI (1170) SrfI (1394) AflII (1416) Bsu36I (1428) BstEII (1497) Bpu10I (1700) PpuMI (2077) BamHI (2607) NgoMIV (2613) NaeI (2615) SgrAI (2619) BstZ17I (2627) EcoRI (2633) HindIII (2752) PaeR7I - PspXI - XhoI (3004) AscI (3368) BspEI (3473) NruI (3832) BfuAI - BspMI (4109) pRetroX-TetOne™ 8508 bp
BspQI  (8267)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (8267)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PciI  (8150)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AlwNI  (7741)
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.
ScaI  (6781)
1 site
A G T A C T T C A T G A
SspI  (6457)
1 site
A A T A T T T T A T A A
SacII  (5120)
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  (5119)
1 site
C A C N N N N G T G G T G N N N N C A C
MluI  (5092)
1 site
A C G C G T T G C G C A
EcoRV  (4734)
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.
PflMI  (4557)
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.
BsiWI  (4433)
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.
AvrII  (77)
1 site
C C T A G G G G A T C C
SnaBI  (519)
1 site
T A C G T A A T G C A T
AfeI  (717)
1 site
A G C G C T T C G C G A
Eco53kI  (733)
1 site
G A G C T C C T C G A G
SacI  (735)
1 site
G A G C T C C T C G A G
PshAI  (1170)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SrfI  (1394)
1 site
G C C C G G G C C G G G C C C G
AflII  (1416)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
Bsu36I  (1428)
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.
BstEII  (1497)
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.
Bpu10I  (1700)
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.
PpuMI  (2077)
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.
BamHI  (2607)
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.
NgoMIV  (2613)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (2615)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
SgrAI  (2619)
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.
BstZ17I  (2627)
1 site
G T A T A C C A T A T G
EcoRI  (2633)
1 site
G A A T T C C T T A A G
HindIII  (2752)
1 site
A A G C T T T T C G A A
PaeR7I  (3004)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3004)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3004)
1 site
C T C G A G G A G C T C
AscI  (3368)
1 site
G G C G C G C C C C G C G C G G
BspEI  (3473)
1 site
T C C G G A A G G C C T
NruI  (3832)
1 site
T C G C G A A G C G C T
BfuAI  (4109)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (4109)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
AmpR
6475 .. 7335  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6475 .. 6543  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6475 .. 7335  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6544 .. 7335  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6475 .. 7335  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CMV/MMLV 5' LTR
164 .. 911  =  748 bp
chimera between the CMV enhancer/promoter and
the 5' LTR of Moloney murine leukemia virus
CMV/MMLV 5' LTR
164 .. 911  =  748 bp
chimera between the CMV enhancer/promoter and
the 5' LTR of Moloney murine leukemia virus
Tet-On® 3G
3552 .. 4298  =  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
3552 .. 4298  =  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
7506 .. 8094  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7506 .. 8094  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
3023 .. 3533  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
3023 .. 3533  =  511 bp
human phosphoglycerate kinase 1 promoter
WPRE
4316 .. 4719  =  404 bp
woodchuck posttranscriptional regulatory element
WPRE
4316 .. 4719  =  404 bp
woodchuck posttranscriptional regulatory element
TRE3GS promoter
2639 .. 3006  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
TRE3GS promoter
2639 .. 3006  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
MMLV Ψ
974 .. 1331  =  358 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
974 .. 1331  =  358 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
3' LTR (ΔU3)
4880 .. 5092  =  213 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
3' LTR (ΔU3)
4880 .. 5092  =  213 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
SV40 poly(A) signal
2298 .. 2432  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2298 .. 2432  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5735 .. 5856  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5735 .. 5856  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6370 .. 6474  =  105 bp
AmpR promoter
6370 .. 6474  =  105 bp
MCS
2607 .. 2638  =  32 bp
multiple cloning site
MCS
2607 .. 2638  =  32 bp
multiple cloning site
tet operator
2764 .. 2782  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2764 .. 2782  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2800 .. 2818  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2800 .. 2818  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2836 .. 2854  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2836 .. 2854  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2872 .. 2890  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2872 .. 2890  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2908 .. 2926  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2908 .. 2926  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2944 .. 2962  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2944 .. 2962  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2980 .. 2998  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2980 .. 2998  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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