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Plasmid Files

pRetroX-Tight-Pur

Retroviral vector for expressing a gene with the Tet-On® Advanced or Tet-Off® Advanced system.

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pRetroX-Tight-Pur Sequence and MappRetroX-Tight-Pur.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BtgZI (278) SspI (6222) XmnI (6017) ScaI (5898) PvuI (5788) FspI (5640) PciI (4525) BspQI - SapI (4409) AvrII (4278) SfiI (4231) AccI (3397) SalI (3396) BmtI (3307) NheI (3303) SnaBI (284) AscI (542) PshAI (676) AfeI (1088) BglII (1576) PspXI (1600) BamHI (1925) NotI (1932) NgoMIV (1939) NaeI (1941) XbaI * (1945) NruI * (1953) MluI (1957) EcoRI (1963) AgeI (2083) BlpI (2246) BsmI (2404) BfuAI - BspMI (2482) BsiWI (2549) RsrII (2609) DraIII (3064) EcoRV (3161) PvuII (3204) pRetroX-Tight-Pur 6567 bp
BtgZI  (278)
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.
SspI  (6222)
1 site
A A T A T T T T A T A A
XmnI  (6017)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5898)
1 site
A G T A C T T C A T G A
PvuI  (5788)
1 site
C G A T C G G C T A G C
FspI  (5640)
1 site
T G C G C A A C G C G T
PciI  (4525)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4409)
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  (4409)
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  (4278)
1 site
C C T A G G G G A T C C
SfiI  (4231)
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  (3397)
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  (3396)
1 site
G T C G A C C A G C T G
BmtI  (3307)
1 site
G C T A G C C G A T C G
NheI  (3303)
1 site
G C T A G C C G A T C G
SnaBI  (284)
1 site
T A C G T A A T G C A T
AscI  (542)
1 site
G G C G C G C C C C G C G C G G
PshAI  (676)
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  (1088)
1 site
A G C G C T T C G C G A
BglII  (1576)
1 site
A G A T C T T C T A G A
PspXI  (1600)
1 site
V C T C G A G B B G A G C T C V
BamHI  (1925)
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.
NotI  (1932)
1 site
G C G G C C G C C G C C G G C G
NgoMIV  (1939)
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  (1941)
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.
XbaI  (1945)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
NruI  (1953)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
MluI  (1957)
1 site
A C G C G T T G C G C A
EcoRI  (1963)
1 site
G A A T T C C T T A A G
AgeI  (2083)
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  (2246)
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  (2404)
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  (2482)
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  (2482)
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  (2549)
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  (2609)
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.
DraIII  (3064)
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  (3161)
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  (3204)
1 site
C A G C T G G T C G A C
Amp
5345 .. 6205  =  861 bp
286 amino acids  =  31.5 kDa
Amp
5345 .. 6205  =  861 bp
286 amino acids  =  31.5 kDa
PuroR
2493 .. 3092  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2493 .. 3092  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4586 .. 5174  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4586 .. 5174  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
PGK promoter
1973 .. 2472  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
1973 .. 2472  =  500 bp
mouse phosphoglycerate kinase 1 promoter
3' LTR (ΔU3)
3273 .. 3698  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
3' LTR (ΔU3)
3273 .. 3698  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
CMV enhancer
6496 .. 308  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
6496 .. 308  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
3964 .. 4293  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3964 .. 4293  =  330 bp
SV40 enhancer and early promoter
tight TRE promoter
1606 .. 1917  =  312 bp
Tet-responsive promoter PTight
tight TRE promoter
1606 .. 1917  =  312 bp
Tet-responsive promoter PTight
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus
(MMLV)
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine
sarcoma virus
AmpR promoter
6206 .. 6310  =  105 bp
AmpR promoter
6206 .. 6310  =  105 bp
MCS
1925 .. 1968  =  44 bp
multiple cloning site
MCS
1925 .. 1968  =  44 bp
multiple cloning site
SV40 ori
4144 .. 4279  =  136 bp
SV40 origin of replication
SV40 ori
4144 .. 4279  =  136 bp
SV40 origin of replication
tet operator
1611 .. 1629  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1611 .. 1629  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1647 .. 1665  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1647 .. 1665  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1682 .. 1700  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1682 .. 1700  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1718 .. 1736  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1718 .. 1736  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1754 .. 1772  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1754 .. 1772  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1789 .. 1807  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1789 .. 1807  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1825 .. 1843  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1825 .. 1843  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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