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pT7Blue-3

Cloning vector with a T7 promoter and dual ampicillin/kanamycin resistance, and with a multiple cloning site that includes an EcoRV site flanked by EcoRI sites.

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pT7Blue-3.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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KpnI (55) Acc65I (51) BspQI - SapI (3597) PciI (3480) PspFI (3180) BseYI (3176) AlwNI (3071) PflMI (2630) Bpu10I (2384) BsmBI (2383) AsiSI (2367) EcoNI (2279) SmaI (2241) TspMI - XmaI (2239) BspDI - ClaI (2058) NruI (2024) SphI (63) PstI (68) MluI (70) SnaBI (78) BamHI (84) EcoRI (91) EcoRV (101) EcoRI (106) SalI (112) AccI (113) HincII (114) HindIII (118) PaeR7I - XhoI (124) AvrII - StyI (130) NheI (135) BmtI (139) XbaI (141) AleI - PmlI (152) BstXI (154) EcoO109I - PspOMI (159) ApaI (163) Eco53kI (167) SacI (169) EagI - NotI (172) AanI - PsiI (439) DraIII (567) BtgZI (568) NgoMIV (668) NaeI (670) BsaHI (1177) TatI (1234) ScaI (1236) NmeAIII (1570) BpmI (1648) BsaI (1651) AhdI (1717) pT7Blue-3 3821 bp
KpnI  (55)
1 site
G G T A C C C C A T G G
Acc65I  (51)
1 site
G G T A C C C C A T G G
BspQI  (3597)
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  (3597)
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.
PciI  (3480)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (3180)
1 site
C C C A G C G G G T C G
BseYI  (3176)
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  (3071)
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.
PflMI  (2630)
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.
Bpu10I  (2384)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsmBI  (2383)
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.
AsiSI  (2367)
1 site
G C G A T C G C C G C T A G C G
EcoNI  (2279)
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.
SmaI  (2241)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (2239)
1 site
C C C G G G G G G C C C
XmaI  (2239)
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.
BspDI  (2058)
1 site
A T C G A T T A G C T A
ClaI  (2058)
1 site
A T C G A T T A G C T A
NruI  (2024)
1 site
T C G C G A A G C G C T
SphI  (63)
1 site
G C A T G C C G T A C G
PstI  (68)
1 site
C T G C A G G A C G T C
MluI  (70)
1 site
A C G C G T T G C G C A
SnaBI  (78)
1 site
T A C G T A A T G C A T
BamHI  (84)
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.
EcoRI  (91)
2 sites
G A A T T C C T T A A G
EcoRV  (101)
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.
EcoRI  (106)
2 sites
G A A T T C C T T A A G
SalI  (112)
1 site
G T C G A C C A G C T G
AccI  (113)
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  (114)
1 site
G T Y R A C C A R Y T G
HindIII  (118)
1 site
A A G C T T T T C G A A
PaeR7I  (124)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (124)
1 site
C T C G A G G A G C T C
AvrII  (130)
1 site
C C T A G G G G A T C C
StyI  (130)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NheI  (135)
1 site
G C T A G C C G A T C G
BmtI  (139)
1 site
G C T A G C C