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Plasmid Files

pQE-81L

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

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pQE-81L Sequence and MappQE-81L.dna
Map and Sequence File   
Sequence Author:  Qiagen
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 T5 promoter PaeR7I - XhoI (1) AatII (4683) ZraI (4681) XmnI (4360) PvuI (4131) BglI (3881) BsaI (3822) AhdI (3761) AlwNI (3284) PciI (2868) BspQI - SapI (2752) NdeI (2691) BstZ17I (2641) PflFI - Tth111I (2615) PfoI (2512) FspAI (2448) PpuMI * (2431) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS BseRI (115) ATG 6xHis BamHI (147) SphI (157) Eco53kI (161) SacI (163) Acc65I (165) KpnI (169) TspMI - XmaI (170) SmaI (172) SalI (175) PstI (185) BfuAI - BspMI (188) HindIII (189) BlpI (201) NheI (309) BmtI (313) Bpu10I (334) BspEI (557) PasI (793) NcoI - StyI (862) KasI (1339) NarI * (1340) SfoI (1341) PluTI (1343) HpaI (1476) EcoRV (1532) BssHII (1567) PspOMI (1771) ApaI (1775) BstEII (1796) BclI * (1964) MluI (1978) pQE-81L 4753 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  (4683)
1 site
G A C G T C C T G C A G
ZraI  (4681)
1 site
G A C G T C C T G C A G
XmnI  (4360)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (4131)
1 site
C G A T C G G C T A G C
BglI  (3881)
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  (3822)
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  (3761)
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  (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.
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.
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
TspMI  (170)
1 site
C C C G G G G G G C C C
XmaI  (170)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (172)
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  (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
Bpu10I  (334)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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  (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.
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.
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).
AmpR
3688 .. 4548  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3688 .. 4479  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3688 .. 4548  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4480 .. 4548  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3688 .. 4548  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
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
AmpR promoter
4549 .. 4653  =  105 bp
AmpR promoter
4549 .. 4653  =  105 bp
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
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