pGEM-T (linearized)

Linearized vector for TA cloning of PCR products. The insertion site is flanked by BstZI sites.

Sequence Author: Promega

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PspOMI (2960) NaeI (2642) NgoMIV (2640) BtgZI (2540) BsaAI - DraIII (2539) PsiI (2411) XmnI (1944) ScaI (1825) TatI (1823) NmeAIII (1493) BpmI (1415) BsaI (1406) ApaI (2964) ZraI (2968) AatII (2970) SphI (2976) BstZI (2981) NcoI - StyI (2987) SfiI (2989) SacII (2996) End (3002) Start (1) SpeI (5) BfuAI - BspMI - BstZI - NotI (12) PstI - SbfI (23) SalI (25) AccI (26) HincII (27) NdeI (32) Eco53kI (42) SacI (44) MluI (49) BstXI (53) NsiI (62) BspQI - SapI (336) PciI (452) BseYI (756) PspFI (760) AlwNI (868) AhdI (1345) pGEM®-T 3001 bp
PspOMI  (2960)
1 site
G G G C C C C C C G G G
NaeI  (2642)
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  (2640)
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  (2540)
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  (2539)
1 site
Y A C G T R R T G C A Y
DraIII  (2539)
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  (2411)
1 site
T T A T A A A A T A T T
XmnI  (1944)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1825)
1 site
A G T A C T T C A T G A
TatI  (1823)
1 site
W G T A C W W C A T G W
NmeAIII  (1493)
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).
BpmI  (1415)
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.
BsaI  (1406)
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.
ApaI  (2964)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
ZraI  (2968)
1 site
G A C G T C C T G C A G
AatII  (2970)
1 site
G A C G T C C T G C A G
SphI  (2976)
1 site
G C A T G C C G T A C G
BstZI  (2981)
2 sites
C G G C C G G C C G G C
NcoI  (2987)
1 site
C C A T G G G G T A C C
StyI  (2987)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
SfiI  (2989)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SacII  (2996)
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.
End  (3002)
0 sites
Start  (1)
0 sites
SpeI  (5)
1 site
A C T A G T T G A T C A
BfuAI  (12)
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  (12)
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  (12)
2 sites
C G G C C G G C C G G C
NotI  (12)
1 site
G C G G C C G C C G C C G G C G
PstI  (23)
1 site
C T G C A G G A C G T C
SbfI  (23)
1 site
C C T G C A G G G G A C G T C C
SalI  (25)
1 site
G T C G A C C A G C T G
AccI  (26)
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  (27)
1 site
G T Y R A C C A R Y T G
NdeI  (32)
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  (42)
1 site
G A G C T C C T C G A G
SacI  (44)
1 site
G A G C T C C T C G A G
MluI  (49)
1 site
A C G C G T T G C G C A
BstXI  (53)
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  (62)
1 site
A T G C A T T A C G T A
BspQI  (336)
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  (336)
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  (452)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (756)
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.
PspFI  (760)
1 site
C C C A G C G G G T C G
AlwNI  (868)
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  (1345)
1 site
G A C N N N N N G T C C T G N N N N N C