pQE-12

Bacterial vector for expressing C-terminally 6xHis-tagged proteins. See pQE-70 for an improved version of this vector.

Sequence Author: Qiagen

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BbsI (3413) EcoO109I (3411) AatII (3357) ZraI (3355) XmnI (3034) PvuI (2805) FspI (2657) AseI (2607) NmeAIII (2583) BglI (2555) BsrFI (2515) BsaI (2496) AhdI (2435) AlwNI (1958) PspFI (1850) BseYI (1846) AvaI - BsoBI - PaeR7I - XhoI (1) T5 promoter PsiI (49) MfeI (59) lac operator EcoRI (88) RBS BseRI (115) ATG BamHI (121) BsaBI * (126) BglII (127) HindIII (152) stop codons BlpI (164) NheI (272) BmtI (276) Bpu10I (297) PvuII (424) BspEI (520) PasI - PflMI * (756) MscI (791) BtgI - NcoI - StyI (825) XbaI (1129) PfoI (1186) PflFI - Tth111I (1289) BsaAI (1296) AccI (1314) BstZ17I (1315) NdeI (1365) BstAPI (1366) BspQI - SapI (1426) AflIII - PciI (1542) pQE-12 3427 bp
BbsI  (3413)
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  (3411)
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  (3357)
1 site
G A C G T C C T G C A G
ZraI  (3355)
1 site
G A C G T C C T G C A G
XmnI  (3034)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (2805)
1 site
C G A T C G G C T A G C
FspI  (2657)
1 site
T G C G C A A C G C G T
AseI  (2607)
1 site
A T T A A T T A A T T A
NmeAIII  (2583)
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  (2555)
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  (2515)
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  (2496)
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  (2435)
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  (1958)
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  (1850)
1 site
C C C A G C G G G T C G
BseYI  (1846)
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.
AvaI  (1)
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  (1)
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  (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
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  (121)
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.
BsaBI  (126)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BglII  (127)
1 site
A G A T C T T C T A G A
HindIII  (152)
1 site
A A G C T T T T C G A A
BlpI  (164)
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  (272)
1 site
G C T A G C C G A T C G
BmtI  (276)
1 site
G C T A G C C G A T C G
Bpu10I  (297)
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  (424)
1 site
C A G C T G G T C G A C
BspEI  (520)
1 site
T C C G G A A G G C C T
PasI  (756)
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  (756)
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  (791)
1 site
T G G C C A A C C G G T
BtgI  (825)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (825)
1 site
C C A T G G G G T A C C
StyI  (825)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
XbaI  (1129)
1 site
T C T A G A A G A T C T
PfoI  (1186)
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.
PflFI  (1289)
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  (1289)
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.
BsaAI  (1296)
1 site
Y A C G T R R T G C A Y
AccI  (1314)
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.
BstZ17I  (1315)
1 site
G T A T A C C A T A T G
NdeI  (1365)
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.
BstAPI  (1366)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BspQI  (1426)
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  (1426)
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.
AflIII  (1542)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (1542)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2362 .. 3222  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   2362 .. 3153  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2362 .. 3222  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   3154 .. 3222  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2362 .. 3222  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
311 .. 970  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
311 .. 970  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1603 .. 2191  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1603 .. 2191  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
3223 .. 3327  =  105 bp
AmpR promoter
3223 .. 3327  =  105 bp
lambda t0 terminator
173 .. 267  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
173 .. 267  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1035 .. 1121  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
1035 .. 1121  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
T5 promoter
10 .. 54  =  45 bp
4 segments
   Segment 1:  
   10 .. 24  =  15 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
   Segment 2:  -35  
   25 .. 30  =  6 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
   Segment 3:  
   31 .. 47  =  17 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
   Segment 4:  -10  
   48 .. 54  =  7 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
ATG
115 .. 117  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
115 .. 117  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
133 .. 150  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
133 .. 150  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
lac operator
62 .. 78  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
62 .. 78  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
stop codons
157 .. 167  =  11 bp
stop codons in all three reading frames
stop codons
157 .. 167  =  11 bp
stop codons in all three reading frames
RBS
101 .. 106  =  6 bp
ribosome binding site
RBS
101 .. 106  =  6 bp
ribosome binding site
ORF:  311 .. 970  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  2492 .. 2758  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2362 .. 3222  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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