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pRetroX-TRE3G

Retroviral vector for doxycycline-inducible expression of a gene in cells containing the Tet-On® 3G transactivator protein.

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pRetroX-TRE3G Sequence and MappRetroX-TRE3G.dna
Map and Sequence File   
Sequence Author:  Clontech
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 CMV enhancer SspI (6255) XmnI (6050) ScaI (5931) PvuI (5821) FspI (5673) PciI (4558) BspQI - SapI (4442) AvrII (4311) SfiI (4264) AccI (3430) SalI (3429) BmtI (3340) BtgZI (279) SnaBI (285) AscI (543) PshAI (677) AfeI (1089) BglII (1577) PspXI (1605) BamHI (1958) BspDI - ClaI (1965) MluI (1970) NotI (1977) MreI - NgoMIV - SgrAI (1984) NaeI (1986) PspOMI (1990) ApaI (1994) EcoRI (1996) AgeI (2116) BlpI (2279) BsmI (2437) BfuAI - BspMI (2515) BsiWI (2582) RsrII (2642) SacII (2740) DraIII (3097) EcoRV (3194) PvuII (3237) NheI (3336) pRetroX-TRE3G 6600 bp
SspI  (6255)
1 site
A A T A T T T T A T A A
XmnI  (6050)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5931)
1 site
A G T A C T T C A T G A
PvuI  (5821)
1 site
C G A T C G G C T A G C
FspI  (5673)
1 site
T G C G C A A C G C G T
PciI  (4558)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4442)
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  (4442)
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.
AvrII  (4311)
1 site
C C T A G G G G A T C C
SfiI  (4264)
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.
AccI  (3430)
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.
SalI  (3429)
1 site
G T C G A C C A G C T G
BmtI  (3340)
1 site
G C T A G C C G A T C G
BtgZI  (279)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SnaBI  (285)
1 site
T A C G T A A T G C A T
AscI  (543)
1 site
G G C G C G C C C C G C G C G G
PshAI  (677)
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.
AfeI  (1089)
1 site
A G C G C T T C G C G A
BglII  (1577)
1 site
A G A T C T T C T A G A
PspXI  (1605)
1 site
V C T C G A G B B G A G C T C V
BamHI  (1958)
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.
BspDI  (1965)
1 site
A T C G A T T A G C T A
ClaI  (1965)
1 site
A T C G A T T A G C T A
MluI  (1970)
1 site
A C G C G T T G C G C A
NotI  (1977)
1 site
G C G G C C G C C G C C G G C G
MreI  (1984)
1 site
C G C C G G C G G C G G C C G C
NgoMIV  (1984)
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.
SgrAI  (1984)
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.
NaeI  (1986)
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.
PspOMI  (1990)
1 site
G G G C C C C C C G G G
ApaI  (1994)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoRI  (1996)
1 site
G A A T T C C T T A A G
AgeI  (2116)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BlpI  (2279)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsmI  (2437)
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.
BfuAI  (2515)
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  (2515)
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.
BsiWI  (2582)
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.
RsrII  (2642)
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.
SacII  (2740)
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.
DraIII  (3097)
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.
EcoRV  (3194)
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.
PvuII  (3237)
1 site
C A G C T G G T C G A C
NheI  (3336)
1 site
G C T A G C C G A T C G
AmpR
5378 .. 6238  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   5378 .. 6169  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5378 .. 6238  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   6170 .. 6238  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5378 .. 6238  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
2526 .. 3125  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2526 .. 3125  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4619 .. 5207  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4619 .. 5207  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
2006 .. 2505  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
2006 .. 2505  =  500 bp
mouse phosphoglycerate kinase 1 promoter
3' LTR (ΔU3)
3306 .. 3731  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
3' LTR (ΔU3)
3306 .. 3731  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
gag (truncated)
1152 .. 1568  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1152 .. 1568  =  417 bp
truncated MMLV gag gene lacking the start codon
CMV enhancer
6529 .. 309  =  381 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
6529 .. 309  =  381 bp
human cytomegalovirus immediate early enhancer
TRE3GV promoter
1611 .. 1957  =  347 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On 3G; optimized for
retroviral and lentiviral vectors
TRE3GV promoter
1611 .. 1957  =  347 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On 3G; optimized for
retroviral and lentiviral vectors
SV40 promoter
3997 .. 4326  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3997 .. 4326  =  330 bp
SV40 enhancer and early promoter
CMV promoter
310 .. 513  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
310 .. 513  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
MMLV Ψ
752 .. 951  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
752 .. 951  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
5' LTR (truncated)
514 .. 689  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
5' LTR (truncated)
514 .. 689  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
AmpR promoter
6239 .. 6343  =  105 bp
AmpR promoter
6239 .. 6343  =  105 bp
MCS
1958 .. 2001  =  44 bp
multiple cloning site
MCS
1958 .. 2001  =  44 bp
multiple cloning site
SV40 ori
4177 .. 4312  =  136 bp
SV40 origin of replication
SV40 ori
4177 .. 4312  =  136 bp
SV40 origin of replication
tet operator
1616 .. 1634  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1616 .. 1634  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1652 .. 1670  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1652 .. 1670  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1688 .. 1706  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1688 .. 1706  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1724 .. 1742  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1724 .. 1742  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1760 .. 1778  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1760 .. 1778  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1796 .. 1814  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1796 .. 1814  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1832 .. 1850  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1832 .. 1850  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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