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Plasmid Files

pTRE2hyg

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

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pTRE2hyg Sequence and MappTRE2hyg.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SV40 promoter SfiI (5073) AvrII (5019) PshAI (4972) BfuAI - BspMI (4682) PstI (4666) AsiSI (4637) RsrII (4588) BtgZI (4323) BsaBI * (3959) BsmI (3872) HpaI (3858) XmnI (3378) FspI (3001) BsaI (2840) AhdI (2779) SexAI * (5251) BamHI (471) PvuII (485) MluI (489) NheI (495) BmtI (499) NotI (502) BspDI - ClaI (510) SalI (521) AccI (522) EcoRV (529) KflI - PpuMI (564) PspOMI (1081) ApaI (1085) Bsu36I (1185) BsgI (1211) MscI (1226) BstXI (1234) BglII (1255) PflMI (1462) BspQI - SapI (1770) pTRE2hyg 5337 bp
SfiI  (5073)
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.
AvrII  (5019)
1 site
C C T A G G G G A T C C
PshAI  (4972)
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.
BfuAI  (4682)
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  (4682)
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.
PstI  (4666)
1 site
C T G C A G G A C G T C
AsiSI  (4637)
1 site
G C G A T C G C C G C T A G C G
RsrII  (4588)
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.
BtgZI  (4323)
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.
BsaBI  (3959)
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.
BsmI  (3872)
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.
HpaI  (3858)
1 site
G T T A A C C A A T T 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
FspI  (3001)
1 site
T G C G C A A C G C G T
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.
SexAI  (5251)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI 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
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.
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).
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
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.
HygR
3975 .. 5000  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
3975 .. 5000  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
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
SV40 promoter
5009 .. 1  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
5009 .. 1  =  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
3777 .. 3858  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3777 .. 3858  =  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
5023 .. 5158  =  136 bp
SV40 origin of replication
SV40 ori
5023 .. 5158  =  136 bp
SV40 origin of replication
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