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Plasmid Files

pT7Blue T-Vector

EcoRV-linearized T7Blue vector with 3-T overhangs and a T7 promoter, for TA cloning of PCR products.

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pT7Blue T-Vector.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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HindIII (2838) BspQI - SapI (2569) AflIII - PciI (2452) PspFI (2152) BseYI (2148) AlwNI (2043) AhdI (1564) BsaI (1498) BpmI (1495) NmeAIII (1417) BfuAI - BspMI (2843) SphI (2848) PstI - SbfI (2854) SalI (2856) AccI (2857) HincII (2858) XbaI (2862) SpeI (2874) NdeI (2881) End (2889) Start (1) BamHI (5) AvaI - BsoBI - TspMI - XmaI (10) BmeT110I (11) SmaI (12) Acc65I (14) KpnI (18) Eco53kI (22) SacI (24) EcoRI (26) PsiI (286) BsaAI - DraIII (414) BtgZI (415) NgoMIV (515) NaeI (517) XmnI (964) BsaHI (1024) TatI (1081) ScaI (1083) pT7Blue T-Vector 2888 bp
HindIII  (2838)
1 site
A A G C T T T T C G A A
BspQI  (2569)
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  (2569)
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.
AflIII  (2452)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2452)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2152)
1 site
C C C A G C G G G T C G
BseYI  (2148)
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  (2043)
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  (1564)
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  (1498)
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  (1495)
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  (1417)
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).
BfuAI  (2843)
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  (2843)
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.
SphI  (2848)
1 site
G C A T G C C G T A C G
PstI  (2854)
1 site
C T G C A G G A C G T C
SbfI  (2854)
1 site
C C T G C A G G G G A C G T C C
SalI  (2856)
1 site
G T C G A C C A G C T G
AccI  (2857)
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  (2858)
1 site
G T Y R A C C A R Y T G
XbaI  (2862)
1 site
T C T A G A A G A T C T
SpeI  (2874)
1 site
A C T A G T T G A T C A
NdeI  (2881)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
End  (2889)
0 sites
Start  (1)
0 sites
BamHI  (5)
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.
AvaI  (10)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (10)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (10)
1 site
C C C G G G G G G C C C
XmaI  (10)
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.
BmeT110I  (11)
1 site
C Y C G R G G R G C Y C
SmaI  (12)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Acc65I  (14)
1 site
G G T A C C C C A T G G
KpnI  (18)
1 site
G G T A C C C C A T G G
Eco53kI  (22)
1 site
G A G C T C C T C G A G
SacI  (24)
1 site
G A G C T C C T C G A G
EcoRI  (26)
1 site
G A A T T C C T T A A G
PsiI  (286)
1 site
T T A T A A A A T A T T
BsaAI  (414)
1 site
Y A C G T R R T G C A Y
DraIII  (414)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BtgZI  (415)
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.
NgoMIV  (515)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (517)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
XmnI  (964)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (1024)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
TatI  (1081)
1 site
W G T A C W W C A T G W
ScaI  (1083)
1 site
A G T A C T T C A T G A
AmpR
777 .. 1637  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   777 .. 845  =  69 bp
   23 amino acids  =  2.6 kDa
AmpR
777 .. 1637  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   846 .. 1637  =  792 bp
   263 amino acids  =  28.9 kDa
AmpR
777 .. 1637  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
ori
1808 .. 2396  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1808 .. 2396  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
190 .. 645  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
190 .. 645  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
672 .. 776  =  105 bp
AmpR promoter
672 .. 776  =  105 bp
LacZα
2794 .. 2888  =  95 bp
31 amino acids  =  3.6 kDa
Product: LacZα fragment of β-galactosidase
LacZα
2794 .. 2888  =  95 bp
31 amino acids  =  3.6 kDa
Product: LacZα fragment of β-galactosidase
lac promoter
2720 .. 2750  =  31 bp
   Segment 1:  -35  
   2720 .. 2725  =  6 bp
promoter for the E. coli lac operon
lac promoter
2720 .. 2750  =  31 bp
   Segment 2:  
   2726 .. 2743  =  18 bp
promoter for the E. coli lac operon
lac promoter
2720 .. 2750  =  31 bp
   Segment 3:  -10  
   2744 .. 2750  =  7 bp
promoter for the E. coli lac operon
lac promoter
2720 .. 2750  =  31 bp
3 segments
promoter for the E. coli lac operon
MCS
2 .. 31  =  30 bp
multiple cloning site
MCS
2 .. 31  =  30 bp
multiple cloning site
M13 fwd
32 .. 48  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
32 .. 48  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
2758 .. 2774  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
2758 .. 2774  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
LacZα
2 .. 239  =  238 bp
78 amino acids  =  8.9 kDa
Product: LacZα fragment of β-galactosidase
LacZα
2 .. 239  =  238 bp
78 amino acids  =  8.9 kDa
Product: LacZα fragment of β-galactosidase
MCS
2838 .. 2888  =  51 bp
multiple cloning site
MCS
2838 .. 2888  =  51 bp
multiple cloning site
T7 promoter
2817 .. 2835  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2817 .. 2835  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
2782 .. 2798  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
2782 .. 2798  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  3 .. 239  =  237 bp
ORF:  78 amino acids  =  8.9 kDa  (no start codon)
ORF:  777 .. 1637  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1241 .. 1507  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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