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Plasmid Files

pT7CFE1-CGST-HA-His

Vector with C-terminal cleavable GST and HA and 6xHis tags, for use in a human in vitro protein expression system.

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pT7CFE1-CGST-HA-His.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Pierce)
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PvuII (4282) PsiI (3849) PfoI (3560) AatII (3449) ZraI (3447) NmeAIII (2675) BpmI (2597) BsaI (2588) AhdI (2527) AvrII (110) HindIII (183) PmlI (275) AarI - BfuAI - BspMI (298) Acc65I (400) KpnI (404) PflMI (412) BtgI (492) MscI (536) XcmI (544) NdeI (546) BstXI (547) BamHI (551) EcoRI (557) EcoRV (565) PacI (573) PstI (582) Eco53kI (586) SacI (588) SalI - SgrDI (590) AccI (591) HincII (592) EagI - NotI (597) AvaI - BsoBI - PaeR7I - PspXI - XhoI (605) BmeT110I (606) EcoNI (642) BspQI - SapI (711) BsgI (918) BstBI (1029) SwaI (1059) BclI * (1066) BglII (1379) PmeI (1423) SpeI (1428) BlpI (1437) BseYI (1938) PspFI (1942) AlwNI (2050) pT7CFE1-CGST-HA-His 4375 bp
PvuII  (4282)
1 site
C A G C T G G T C G A C
PsiI  (3849)
1 site
T T A T A A A A T A T T
PfoI  (3560)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AatII  (3449)
1 site
G A C G T C C T G C A G
ZraI  (3447)
1 site
G A C G T C C T G C A G
NmeAIII  (2675)
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  (2597)
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  (2588)
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  (2527)
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.
AvrII  (110)
1 site
C C T A G G G G A T C C
HindIII  (183)
1 site
A A G C T T T T C G A A
PmlI  (275)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (298)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (298)
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  (298)
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.
Acc65I  (400)
1 site
G G T A C C C C A T G G
KpnI  (404)
1 site
G G T A C C C C A T G G
PflMI  (412)
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.
BtgI  (492)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
MscI  (536)
1 site
T G G C C A A C C G G T
XcmI  (544)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
NdeI  (546)
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.
BstXI  (547)
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.
BamHI  (551)
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  (557)
1 site
G A A T T C C T T A A G
EcoRV  (565)
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.
PacI  (573)
1 site
T T A A T T A A A A T T A A T T
PstI  (582)
1 site
C T G C A G G A C G T C
Eco53kI  (586)
1 site
G A G C T C C T C G A G
SacI  (588)
1 site
G A G C T C C T C G A G
SalI  (590)
1 site
G T C G A C C A G C T G
SgrDI  (590)
1 site
C G T C G A C G G C A G C T G C
AccI  (591)
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  (592)
1 site
G T Y R A C C A R Y T G
EagI  (597)
1 site
C G G C C G G C C G G C
NotI  (597)
1 site
G C G G C C G C C G C C G G C G
AvaI  (605)
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  (605)
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.
PaeR7I  (605)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (605)
1 site
V C T C G A G B B G A G C T C V
XhoI  (605)
1 site
C T C G A G G A G C T C
BmeT110I  (606)
1 site
C Y C G R G G R G C Y C
EcoNI  (642)
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.
BspQI  (711)
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  (711)
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.
BsgI  (918)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BstBI  (1029)
1 site
T T C G A A A A G C T T
SwaI  (1059)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BclI  (1066)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BglII  (1379)
1 site
A G A T C T T C T A G A
PmeI  (1423)
1 site
G T T T A A A C C A A A T T T G
SpeI  (1428)
1 site
A C T A G T T G A T C A
BlpI  (1437)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BseYI  (1938)
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  (1942)
1 site
C C C A G C G G G T C G
AlwNI  (2050)
1 site
C A G N N N C