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Plasmid Files

pTRE2

Expression vector with a Tet-responsive promoter.

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pTRE2 Sequence and MappTRE2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 AatII (3701) ZraI (3699) XmnI (3378) PvuI (3149) FspI (3001) NmeAIII (2927) BsrFI (2859) BsaI (2840) AhdI (2779) PspFI (2194) BseYI (2190) DrdI (1994) PaeR7I - PspXI - XhoI (1) minimal CMV promoter StuI (339) BsmBI (377) SacII (443) BamHI (471) PvuII (485) MCS MluI (489) NheI (495) BmtI (499) EagI - NotI (502) BspDI - ClaI (510) HindIII (515) SalI (521) AccI (522) EcoRV (529) XbaI (533) KflI - PpuMI (564) NcoI - StyI (670) MfeI (906) PsiI (914) PspOMI (1081) ApaI (1085) Bsu36I (1185) BsgI (1211) MscI (1226) BstXI (1234) BglII (1255) BstAPI (1451) PflMI (1462) BspQI - SapI (1770) pTRE2 3770 bp
AatII  (3701)
1 site
G A C G T C C T G C A G
ZraI  (3699)
1 site
G A C G T C C T G C A G
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
NmeAIII  (2927)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrFI  (2859)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
BsaI  (2840)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
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.
PspFI  (2194)
1 site
C C C A G C G G G T C G
BseYI  (2190)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
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.
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1)
1 site
C T C G A G G A G C T C
StuI  (339)
1 site
A G G C C T T C C G G A
BsmBI  (377)
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.
SacII  (443)
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.
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
EagI  (502)
1 site
C G G C C G G C C G G C
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
HindIII  (515)
1 site
A A G C T T T T C G A A
SalI  (521)
1 site
G T C G A C C A G C T G
AccI  (522)
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.
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.
XbaI  (533)
1 site
T C T A G A A G A T C T
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.
NcoI  (670)
1 site
C C A T G G G G T A C C
StyI  (670)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
MfeI  (906)
1 site
C A A T T G G T T A A C
PsiI  (914)
1 site
T T A T A A A A T A T T
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).
MscI  (1226)
1 site
T G G C C A A C C G G T
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
BstAPI  (1451)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
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.
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
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
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
MCS
440 .. 538  =  99 bp
multiple cloning site
MCS
440 .. 538  =  99 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
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