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Plasmid Files

pTA Plus (linearized)

Linearized TA cloning vector with 3'-T overhangs derived from pPCR-Script Amp.

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pTA Plus (linearized) Sequence and MappTA Plus (linearized).dna
Map and Sequence File   
Sequence Author:  EMBL
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 AanI - PsiI (615) DraIII (490) BsaAI (487) NaeI (384) NgoMIV (382) M13 fwd KpnI (61) Acc65I (57) ApaI (55) EcoO109I - PspOMI (51) AbsI - PaeR7I - PspXI - XhoI (42) HincII (38) AccI (37) SalI (36) BspDI - ClaI (28) HindIII (21) EcoRV (17) EcoRI (9) < XcmI > (1) < XcmI > (2965) SmaI - SrfI (2947) TspMI - XmaI (2945) EagI - NotI (2933) BstXI (2928) SacII (2927) BtgI (2924) SacI (2920) Eco53kI (2918) M13 rev BspQI - SapI (2635) NspI (2522) AflIII - PciI (2518) PspFI (2218) XmnI (1030) BsaHI (1090) ScaI (1149) BsaI (1564) AhdI (1630) AlwNI (2109) BseYI (2214) pTA Plus 2964 bp
AanI  (615)
1 site
T T A T A A A A T A T T
PsiI  (615)
1 site
T T A T A A A A T A T T
DraIII  (490)
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.
BsaAI  (487)
1 site
Y A C G T R R T G C A Y
NaeI  (384)
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.
NgoMIV  (382)
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.
KpnI  (61)
1 site
G G T A C C C C A T G G
Acc65I  (57)
1 site
G G T A C C C C A T G G
ApaI  (55)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (51)
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  (51)
1 site
G G G C C C C C C G G G
AbsI  (42)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (42)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (42)
1 site
V C T C G A G B B G A G C T C V
XhoI  (42)
1 site
C T C G A G G A G C T C
HincII  (38)
1 site
G T Y R A C C A R Y T G
AccI  (37)
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.
SalI  (36)
1 site
G T C G A C C A G C T G
BspDI  (28)
1 site
A T C G A T T A G C T A
ClaI  (28)
1 site
A T C G A T T A G C T A
HindIII  (21)
1 site
A A G C T T T T C G A A
EcoRV  (17)
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  (9)
1 site
G A A T T C C T T A A G
Start  (1)
0 sites
End  (2965)
0 sites
SmaI  (2947)
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  (2947)
1 site
G C C C G G G C C G G G C C C G
TspMI  (2945)
1 site
C C C G G G G G G C C C
XmaI  (2945)
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.
EagI  (2933)
1 site
C G G C C G G C C G G C
NotI  (2933)
1 site
G C G G C C G C C G C C G G C G
BstXI  (2928)
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.
SacII  (2927)
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.
BtgI  (2924)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
SacI  (2920)
1 site
G A G C T C C T C G A G
Eco53kI  (2918)
1 site
G A G C T C C T C G A G
BspQI  (2635)
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  (2635)
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  (2522)
1 site
R C A T G Y Y G T A C R
AflIII  (2518)
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  (2518)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (2218)
1 site
C C C A G C G G G T C G
XmnI  (1030)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaHI  (1090)
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.
ScaI  (1149)
1 site
A G T A C T T C A T G A
BsaI  (1564)
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  (1630)
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.
AlwNI  (2109)
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.
BseYI  (2214)
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.
AmpR
843 .. 1703  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   843 .. 911  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
843 .. 1703  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   912 .. 1703  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
843 .. 1703  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1874 .. 2462  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1874 .. 2462  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
257 .. 712  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
257 .. 712  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
AmpR promoter
738 .. 842  =  105 bp
AmpR promoter
738 .. 842  =  105 bp
lac promoter
2786 .. 2816  =  31 bp
   Segment 1:  -35  
   2786 .. 2791  =  6 bp
promoter for the E. coli lac operon
lac promoter
2786 .. 2816  =  31 bp
   Segment 2:  
   2792 .. 2809  =  18 bp
promoter for the E. coli lac operon
lac promoter
2786 .. 2816  =  31 bp
   Segment 3:  -10  
   2810 .. 2816  =  7 bp
promoter for the E. coli lac operon
lac promoter
2786 .. 2816  =  31 bp
3 segments
promoter for the E. coli lac operon
T7 promoter
71 .. 89  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
71 .. 89  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T3 promoter
2885 .. 2903  =  19 bp
promoter for bacteriophage T3 RNA polymerase
T3 promoter
2885 .. 2903  =  19 bp
promoter for bacteriophage T3 RNA polymerase
KS primer
29 .. 45  =  17 bp
common sequencing primer, one of multiple similar
variants
KS primer
29 .. 45  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
99 .. 115  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
99 .. 115  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
2824 .. 2840  =  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
2824 .. 2840  =  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).
M13 rev
2848 .. 2864  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2848 .. 2864  =  17 bp
common sequencing primer, one of multiple similar
variants
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