pQE-82L

Bacterial lacIq vector for expressing N-terminally 6xHis-tagged proteins. For other reading frames, use pQE-80L or pQE-81L.

Sequence Author: Qiagen

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T5 promoter PaeR7I - XhoI (1) AatII (4682) ZraI (4680) XmnI (4359) PvuI (4130) BglI (3880) BsaI (3821) AhdI (3760) AlwNI (3283) PciI (2867) BspQI - SapI (2751) NdeI (2690) BstZ17I (2640) PflFI - Tth111I (2614) PfoI (2511) FspAI (2447) PpuMI * (2430) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS BseRI (115) ATG 6xHis BamHI (146) SphI (156) Eco53kI (160) SacI (162) Acc65I (164) KpnI (168) TspMI - XmaI (169) SmaI (171) SalI (174) PstI (184) BfuAI - BspMI (187) HindIII (188) BlpI (200) NheI (308) BmtI (312) Bpu10I (333) BspEI (556) PasI (792) KasI (1338) NarI * (1339) SfoI (1340) PluTI (1342) HpaI (1475) EcoRV (1531) BssHII (1566) PspOMI (1770) ApaI (1774) BstEII (1795) BclI * (1963) MluI (1977) pQE-82L 4752 bp
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
AatII  (4682)
1 site
G A C G T C C T G C A G
ZraI  (4680)
1 site
G A C G T C C T G C A G
XmnI  (4359)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (4130)
1 site
C G A T C G G C T A G C
BglI  (3880)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BsaI  (3821)
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  (3760)
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  (3283)
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.
PciI  (2867)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2751)
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  (2751)
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.
NdeI  (2690)
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.
BstZ17I  (2640)
1 site
G T A T A C C A T A T G
PflFI  (2614)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2614)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PfoI  (2511)
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.
FspAI  (2447)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (2430)
1 site
R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
PsiI  (49)
1 site
T T A T A A A A T A T T
MfeI  (59)
1 site
C A A T T G G T T A A C
EcoRI  (88)
1 site
G A A T T C C T T A A G
BseRI  (115)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BamHI  (146)
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.
SphI  (156)
1 site
G C A T G C C G T A C G
Eco53kI  (160)
1 site
G A G C T C C T C G A G
SacI  (162)
1 site
G A G C T C C T C G A G
Acc65I  (164)
1 site
G G T A C C C C A T G G
KpnI  (168)
1 site
G G T A C C C C A T G G
TspMI  (169)
1 site
C C C G G G G G G C C C
XmaI  (169)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (171)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (174)
1 site
G T C G A C C A G C T G
PstI  (184)
1 site
C T G C A G G A C G T C
BfuAI  (187)
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  (187)
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.
HindIII  (188)
1 site
A A G C T T T T C G A A
BlpI  (200)
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.
NheI  (308)
1 site
G C T A G C C G A T C G
BmtI  (312)
1 site
G C T A G C C G A T C G
Bpu10I  (333)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BspEI  (556)
1 site