pSpark TA Done

Linearized vector with 3'-T overhangs and an enhanced multiple cloning site for TA cloning of PCR products.

Sequence Author: Canvax Biotech

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End (3017) EcoRI (3008) SacII (3005) BstZI - NotI (2999) NcoI - StyI (2993) BstZI (2987) SphI (2982) AatII (2976) ZraI (2974) ApaI (2970) PspOMI (2966) NaeI (2648) NgoMIV (2646) BtgZI (2546) BsaAI - DraIII (2545) PsiI (2417) XmnI (1950) ScaI (1831) TatI (1829) Start (1) SpeI (5) EcoRI (11) BfuAI - BspMI - BstZI - NotI (18) PstI - SbfI (29) SalI (31) AccI (32) HincII (33) NdeI (38) Eco53kI (48) SacI (50) MluI (55) BstXI (59) NsiI (68) BspQI - SapI (342) PciI (458) BseYI (762) PspFI (766) AlwNI (874) AhdI (1351) BsaI (1412) BpmI (1421) NmeAIII (1499) pSpark® TA Done 3016 bp
End  (3017)
0 sites
EcoRI  (3008)
2 sites
G A A T T C C T T A A G
SacII  (3005)
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.
BstZI  (2999)
3 sites
C G G C C G G C C G G C
NotI  (2999)
2 sites
G C G G C C G C C G C C G G C G
NcoI  (2993)
1 site
C C A T G G G G T A C C
StyI  (2993)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BstZI  (2987)
3 sites
C G G C C G G C C G G C
SphI  (2982)
1 site
G C A T G C C G T A C G
AatII  (2976)
1 site
G A C G T C C T G C A G
ZraI  (2974)
1 site
G A C G T C C T G C A G
ApaI  (2970)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (2966)
1 site
G G G C C C C C C G G G
NaeI  (2648)
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  (2646)
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.
BtgZI  (2546)
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.
BsaAI  (2545)
1 site
Y A C G T R R T G C A Y
DraIII  (2545)
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.
PsiI  (2417)
1 site
T T A T A A A A T A T T
XmnI  (1950)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1831)
1 site
A G T A C T T C A T G A
TatI  (1829)
1 site
W G T A C W W C A T G W
Start  (1)
0 sites
SpeI  (5)
1 site
A C T A G T T G A T C A
EcoRI  (11)
2 sites
G A A T T C C T T A A G
BfuAI  (18)
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  (18)
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.
BstZI  (18)
3 sites
C G G C C G G C C G G C
NotI  (18)
2 sites
G C G G C C G C C G C C G G C G
PstI  (29)
1 site
C T G C A G G A C G T C
SbfI  (29)
1 site
C C T G C A G G G G A C G T C C
SalI  (31)
1 site
G T C G A C C A G C T G
AccI  (32)
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  (33)
1 site
G T Y R A C C A R Y T G
NdeI  (38)
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.
Eco53kI  (48)
1 site
G A G C T C C T C G A G
SacI  (50)
1 site
G A G C T C C T C G A G
MluI  (55)
1 site
A C G C G T T G C G C A
BstXI  (59)
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.
NsiI  (68)
1 site
A T G C A T T A C G T A
BspQI  (342)
1 site
G C T C T T C N C G A G A A G N N N