pQE-30 UA (linearized)

Linearized bacterial expression and 6xHis-tagging vector with 3'-U overhangs for UA cloning of a PCR product.

Sequence Author: Qiagen

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BbsI (3316) EcoO109I (3314) AatII (3260) ZraI (3258) XmnI (2937) PvuI (2708) FspI (2560) AseI (2510) NmeAIII (2486) BglI (2458) BsrFI (2418) BsaI (2399) AhdI (2338) AlwNI (1861) PspFI (1753) BseYI (1749) PaeR7I - XhoI (3331) T5 promoter PsiI (3379) MfeI (3389) lac operator EcoRI (3418) ATG 6xHis BamHI (3475) PmlI (3483) EcoRV (3488) End (3504) Start (1) BglII (15) Eco53kI (23) BanII - SacI (25) Acc65I (27) KpnI - TspMI - XmaI (31) SmaI (33) SalI (36) HincII (38) StuI (44) PstI (52) HindIII (56) BlpI (68) NheI (176) BmtI (180) Bpu10I (201) PvuII (328) BspEI (424) PasI - PflMI * (660) MscI (695) BtgI - NcoI - StyI (729) XbaI (1032) PfoI (1089) PflFI - Tth111I (1192) BstZ17I (1218) NdeI (1268) BstAPI (1269) BspQI - SapI (1329) AflIII - PciI (1445) pQE-30 UA 3503 bp
BbsI  (3316)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
EcoO109I  (3314)
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.
AatII  (3260)
1 site
G A C G T C C T G C A G
ZraI  (3258)
1 site
G A C G T C C T G C A G
XmnI  (2937)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (2708)
1 site
C G A T C G G C T A G C
FspI  (2560)
1 site
T G C G C A A C G C G T
AseI  (2510)
1 site
A T T A A T T A A T T A
NmeAIII  (2486)
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).
BglI  (2458)
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.
BsrFI  (2418)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BsaI  (2399)
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  (2338)
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  (1861)
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.
PspFI  (1753)
1 site
C C C A G C G G G T C G
BseYI  (1749)
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.
PaeR7I  (3331)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3331)
1 site
C T C G A G G A G C T C
PsiI  (3379)
1 site
T T A T A A A A T A T T
MfeI  (3389)
1 site
C A A T T G G T T A A C
EcoRI  (3418)
1 site
G A A T T C C T T A A G
BamHI  (3475)
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.
PmlI  (3483)
1 site
C A C G T G G T G C A C
EcoRV  (3488)
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.
End  (3504)
0 sites
Start  (1)
0 sites
BglII  (15)
1 site
A G A T C T T C T A G A
Eco53kI  (23)
1 site
G A G C T C C T C G A G
BanII  (25)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (25)
1 site
G A G C T C C T C G A G
Acc65I  (27)
1 site
G G T A C C C C A T G G
KpnI  (31)
1 site
G G T A C C C C A T G G
TspMI  (31)
1 site
C C C G G G G G G C C C
XmaI  (31)
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  (33)
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  (36)
1 site
G T C G A C C A G C T G
HincII  (38)
1 site
G T Y R A C C A R Y T G
StuI  (44)
1 site
A G G C C T T C C G G A
PstI  (52)
1 site
C T G C A G G A C G T C
HindIII  (56)
1 site
A A G C T T T T C G A A
BlpI  (68)
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  (176)
1 site
G C T A G C C G A T C G
BmtI  (180)
1 site
G C T A G C C G A T C G
Bpu10I  (201)
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.
PvuII  (328)
1 site
C A G C T G G T C G A C
BspEI  (424)
1 site
T C C G G A A G G C C T
PasI  (660)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (660)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
MscI  (695)
1 site
T G G C C A A C