Resources
Plasmid Files

pQE-81L-Kan

Bacterial lacIq vector with a kanamycin resistance marker for expressing N-terminally 6xHis-tagged proteins. For other reading frames, use pQE‑80L-Kan or pQE‑82L-Kan.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pQE-81L-Kan Sequence and MappQE-81L-Kan.dna
Map and Sequence File   
Sequence Author:  Qiagen
Download Free Trial Get SnapGene Viewer

 NruI (4350) BspDI - ClaI (4314) EcoNI (4094) AsiSI - PvuI (4009) XmnI (3618) AcuI (3416) AlwNI (3284) PciI (2868) BspQI - SapI (2752) NdeI (2691) BstZ17I (2641) PflFI - Tth111I (2615) PfoI (2512) FspI - FspAI (2448) PpuMI * (2431) PaeR7I - XhoI (1) PsiI (49) MfeI (59) lac operator EcoRI (88) BseRI (115) ATG 6xHis BamHI (147) SphI (157) Eco53kI (161) SacI (163) Acc65I (165) KpnI (169) SalI (175) PstI (185) BfuAI - BspMI (188) HindIII (189) BlpI (201) NheI (309) BmtI (313) BspEI (557) PasI (793) NcoI - StyI (862) ScaI (978) KasI (1339) NarI * (1340) SfoI (1341) PluTI (1343) HpaI (1476) EcoRV (1532) BssHII (1567) PspOMI (1771) ApaI (1775) NmeAIII (1778) BstEII (1796) BclI * (1964) MluI (1978) pQE-81L-Kan 4543 bp
NruI  (4350)
1 site
T C G C G A A G C G C T
BspDI  (4314)
1 site
A T C G A T T A G C T A
ClaI  (4314)
1 site
A T C G A T T A G C T A
EcoNI  (4094)
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.
AsiSI  (4009)
1 site
G C G A T C G C C G C T A G C G
PvuI  (4009)
1 site
C G A T C G G C T A G C
XmnI  (3618)
1 site
G A A N N N N T T C C T T N N N N A A G
AcuI  (3416)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (3284)
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  (2868)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2752)
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  (2752)
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  (2691)
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  (2641)
1 site
G T A T A C C A T A T G
PflFI  (2615)
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  (2615)
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  (2512)
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.
FspI  (2448)
1 site
T G C G C A A C G C G T
FspAI  (2448)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (2431)
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.
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  (147)
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  (157)
1 site
G C A T G C C G T A C G
Eco53kI  (161)
1 site
G A G C T C C T C G A G
SacI  (163)
1 site
G A G C T C C T C G A G
Acc65I  (165)
1 site
G G T A C C C C A T G G
KpnI  (169)
1 site
G G T A C C C C A T G G
SalI  (175)
1 site
G T C G A C C A G C T G
PstI  (185)
1 site
C T G C A G G A C G T C
BfuAI  (188)
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  (188)
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  (189)
1 site
A A G C T T T T C G A A
BlpI  (201)
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  (309)
1 site
G C T A G C C G A T C G
BmtI  (313)
1 site
G C T A G C C G A T C G
BspEI  (557)
1 site
T C C G G A A G G C C T
PasI  (793)
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.
NcoI  (862)
1 site
C C A T G G G G T A C C
StyI  (862)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
ScaI  (978)
1 site
A G T A C T T C A T G A
KasI  (1339)
1 site
G G C G C C C C G C G G
NarI  (1340)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (1341)
1 site
G G C G C C C C G C G G
PluTI  (1343)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
HpaI  (1476)
1 site
G T T A A C C A A T T G
EcoRV  (1532)
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.
BssHII  (1567)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
PspOMI  (1771)
1 site
G G G C C C C C C G G G
ApaI  (1775)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NmeAIII  (1778)
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).
BstEII  (1796)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BclI  (1964)
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.
MluI  (1978)
1 site
A C G C G T T G C G C A
lacI
1251 .. 2333  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
1251 .. 2333  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
KanR
3624 .. 4439  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
3624 .. 4439  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
CmR
348 .. 1007  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
348 .. 1007  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2929 .. 3517  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2929 .. 3517  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lambda t0 terminator
210 .. 304  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
210 .. 304  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1072 .. 1158  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
1072 .. 1158  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
lacIq promoter
2334 .. 2411  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
lacIq promoter
2334 .. 2411  =  78 bp
In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about
10-fold.
MCS
147 .. 194  =  48 bp
multiple cloning site
MCS
147 .. 194  =  48 bp
multiple cloning site
T5 promoter
10 .. 54  =  45 bp
   Segment 1:  
   10 .. 24  =  15 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   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
   Segment 3:  
   31 .. 47  =  17 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   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
6xHis
127 .. 144  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
127 .. 144  =  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).
RBS
101 .. 106  =  6 bp
ribosome binding site
RBS
101 .. 106  =  6 bp
ribosome binding site
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
lac operator
30 .. 46  =  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
30 .. 46  =  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
194 .. 204  =  11 bp
stop codons in all three reading frames
stop codons
194 .. 204  =  11 bp
stop codons in all three reading frames
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter