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pT7Blue-2

Multi-purpose cloning vector with T7 and SP6 promoters, and with a multiple cloning site that encodes a cleavable N-terminal S-tag.

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pT7Blue-2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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BspQI - SapI (2879) NspI (2766) PciI (2762) PspFI (2462) BseYI (2458) AlwNI (2353) AhdI (1874) BsaI (1808) BpmI (1805) NmeAIII (1727) MscI (94) BtgI - NcoI (98) BstBI (134) Acc65I (157) AgeI (160) KpnI (161) BseRI - TspMI - XmaI (189) SmaI - SrfI (191) PstI (212) MluI (214) SnaBI (222) BamHI (228) EcoNI (236) EcoRI (253) EcoRV (262) SalI (268) AccI (269) HincII (270) HindIII (274) PaeR7I - XhoI (280) AvrII (286) NheI (291) BmtI (295) XbaI (297) AleI - PmlI (308) BstXI (310) EcoO109I - PspOMI (315) ApaI (319) Eco53kI (323) SacI (325) EagI - NotI (328) PsiI (596) DraIII (724) NgoMIV (825) NaeI (827) XmnI (1274) BsaHI (1334) TatI (1391) ScaI (1393) pT7Blue-2 3103 bp
BspQI  (2879)
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  (2879)
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.
NspI  (2766)
1 site
R C A T G Y Y G T A C R
PciI  (2762)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2462)
1 site
C C C A G C G G G T C G
BseYI  (2458)
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.
AlwNI  (2353)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (1874)
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.
BsaI  (1808)
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.
BpmI  (1805)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (1727)
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).
MscI  (94)
1 site
T G G C C A A C C G G T
BtgI  (98)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (98)
1 site
C C A T G G G G T A C C
BstBI  (134)
1 site
T T C G A A A A G C T T
Acc65I  (157)
1 site
G G T A C C C C A T G G
AgeI  (160)
1 site
A C C G G T T G G C C A
KpnI  (161)
1 site
G G T A C C C C A T G G
BseRI  (189)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
TspMI  (189)
1 site
C C C G G G G G G C C C
XmaI  (189)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (191)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (191)
1 site
G C C C G G G C C G G G C C C G
PstI  (212)
1 site
C T G C A G G A C G T C
MluI  (214)
1 site
A C G C G T T G C G C A
SnaBI  (222)
1 site
T A C G T A A T G C A T
BamHI  (228)
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.
EcoNI  (236)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
EcoRI  (253)
1 site
G A A T T C C T T A A G
EcoRV  (262)
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.
SalI  (268)
1 site
G T C G A C C A G C T G
AccI  (269)
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.
HincII  (270)
1 site
G T Y R A C C A R Y T G
HindIII  (274)
1 site
A A G C T T T T C G A A
PaeR7I  (280)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (280)
1 site
C T C G A G G A G C T C
AvrII  (286)
1 site
C C T A G G G G A T C C
NheI  (291)
1 site
G C T A G C C G A T C G
BmtI  (295)
1 site
G C T A G C C G A T C G
XbaI  (297)
1 site
T C T A G A A G A T C T
AleI  (308)
1 site
C A C N N N N G T G G T G N N N N C A C
PmlI  (308)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BstXI  (310)
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.
EcoO109I  (315)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (315)
1 site
G G G C C C C C C G G G
ApaI  (319)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
Eco53kI  (323)
1 site
G A G C T C C T C G A G
SacI  (325)
1 site
G A G C T C C T C G A G
EagI  (328)
1 site
C G G C C G G C C G G C
NotI  (328)
1 site
G C G G C C G C C G C