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Plasmid Files

pTRE2pur

Expression vector with a Tet-responsive promoter and a puromycin resistance marker.

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pTRE2pur Sequence and MappTRE2pur.dna
Map and Sequence File   
Sequence Author:  Clontech
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 HpaI (5026) BsmI (5014) BsaBI * (4925) DraIII (4769) PfoI * (4673) BssHII (4653) BstEII (4332) RsrII (4314) BsiWI (4254) PflFI - Tth111I (4240) AgeI (4137) PstI - SbfI (4122) BfuAI - BspMI (4111) AvrII (4090) SfiI (4043) AleI (3773) XmnI (3378) PvuI (3149) FspI (3001) AhdI (2779) SV40 poly(A) signal BamHI (471) PvuII (485) MluI (489) NheI (495) BmtI (499) NotI (502) BspDI - ClaI (510) EcoRV (529) KflI - PpuMI (564) PspOMI (1081) ApaI (1085) Bsu36I (1185) BsgI (1211) BstXI (1234) BglII (1255) PflMI (1462) BspQI - SapI (1770) DrdI (1994) pTRE2pur 5108 bp
HpaI  (5026)
1 site
G T T A A C C A A T T G
BsmI  (5014)
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.
BsaBI  (4925)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
DraIII  (4769)
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.
PfoI  (4673)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BssHII  (4653)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BstEII  (4332)
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.
RsrII  (4314)
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.
BsiWI  (4254)
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.
PflFI  (4240)
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  (4240)
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.
AgeI  (4137)
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.
PstI  (4122)
1 site
C T G C A G G A C G T C
SbfI  (4122)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (4111)
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  (4111)
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.
AvrII  (4090)
1 site
C C T A G G G G A T C C
SfiI  (4043)
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.
AleI  (3773)
1 site
C A C N N N N G T G G T G N N N N C A C
XmnI  (3378)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3149)
1 site
C G A T C G G C T A G C
FspI  (3001)
1 site
T G C G C A A C G C G T
AhdI  (2779)
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.
BamHI  (471)
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.
PvuII  (485)
1 site
C A G C T G G T C G A C
MluI  (489)
1 site
A C G C G T T G C G C A
NheI  (495)
1 site
G C T A G C C G A T C G
BmtI  (499)
1 site
G C T A G C C G A T C G
NotI  (502)
1 site
G C G G C C G C C G C C G G C G
BspDI  (510)
1 site
A T C G A T T A G C T A
ClaI  (510)
1 site
A T C G A T T A G C T A
EcoRV  (529)
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.
KflI  (564)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (564)
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.
PspOMI  (1081)
1 site
G G G C C C C C C G G G
ApaI  (1085)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
Bsu36I  (1185)
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.
BsgI  (1211)
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).
BstXI  (1234)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BglII  (1255)
1 site
A G A T C T T C T A G A
PflMI  (1462)
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.
BspQI  (1770)
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  (1770)
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.
DrdI  (1994)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AmpR
2706 .. 3566  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2706 .. 3497  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2706 .. 3566  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3498 .. 3566  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2706 .. 3566  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
4198 .. 4797  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4198 .. 4797  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
1947 .. 2535  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1947 .. 2535  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
β-globin intron
554 .. 1126  =  573 bp
intron from rabbit β-globin gene
β-globin intron
554 .. 1126  =  573 bp
intron from rabbit β-globin gene
SV40 promoter
3776 .. 4105  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3776 .. 4105  =  330 bp
SV40 enhancer and early promoter
tetracycline response element
15 .. 285  =  271 bp
   Segment 1:  tetO  
   15 .. 33  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 2:  
   34 .. 56  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 3:  tetO  
   57 .. 75  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 4:  
   76 .. 98  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 5:  tetO  
   99 .. 117  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 6:  
   118 .. 140  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 7:  tetO  
   141 .. 159  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 8:  
   160 .. 182  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 9:  tetO  
   183 .. 201  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 10:  
   202 .. 224  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 11:  tetO  
   225 .. 243  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 12:  
   244 .. 266  =  23 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
   Segment 13:  tetO  
   267 .. 285  =  19 bp
contains seven copies of the tetracycline operator
tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
contains seven copies of the tetracycline operator
tetO
AmpR promoter
3567 .. 3671  =  105 bp
AmpR promoter
3567 .. 3671  =  105 bp
SV40 poly(A) signal
5027 .. 5108  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5027 .. 5108  =  82 bp
SV40 polyadenylation signal
MCS
471 .. 532  =  62 bp
multiple cloning site
MCS
471 .. 532  =  62 bp
multiple cloning site
β-globin poly(A) signal
1323 .. 1378  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
1323 .. 1378  =  56 bp
rabbit β-globin polyadenylation signal
minimal CMV promoter
318 .. 356  =  39 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
318 .. 356  =  39 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 ori
3956 .. 4091  =  136 bp
SV40 origin of replication
SV40 ori
3956 .. 4091  =  136 bp
SV40 origin of replication
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