pTRE2

Expression vector with a Tet-responsive promoter.

Sequence Author: Clontech (TaKaRa)

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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 - Esp3I (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

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
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.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (377)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I 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
2 segments
   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
2 segments
   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
13 segments
   Segment 1:  tetO  
   15 .. 33  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 2:  
   34 .. 56  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 3:  tetO  
   57 .. 75  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 4:  
   76 .. 98  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 5:  tetO  
   99 .. 117  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 6:  
   118 .. 140  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 7:  tetO  
   141 .. 159  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 8:  
   160 .. 182  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 9:  tetO  
   183 .. 201  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 10:  
   202 .. 224  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 11:  tetO  
   225 .. 243  =  19 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   Segment 12:  
   244 .. 266  =  23 bp
contains seven copies of the tetracycline operator tetO
tetracycline response element
15 .. 285  =  271 bp
13 segments
   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
ORF:  2836 .. 3102  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2706 .. 3566  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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