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Plasmid Files

pQE-16

Bacterial vector for expressing DHFR fusion proteins with a C-terminal 6xHis tag. Similar to pQE‑19 and pQE‑22, but with a different reading frame after the DHFR.

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pQE-16 Sequence and MappQE-16.dna
Map and Sequence File   
Sequence Author:  Qiagen
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 BbsI (3982) EcoO109I (3980) AatII (3926) ZraI (3924) FspI (3226) NmeAIII (3152) BglI (3124) BsrFI (3084) AhdI (3004) PspFI (2419) BseYI (2415) PciI (2111) BspQI - SapI (1995) AvaI - BsoBI - PaeR7I - XhoI (1) BmeT110I (2) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS ATG BamHI (121) SexAI * (301) Eco53kI (391) BanII - SacI (393) BstXI (411) AflII (423) EcoNI (634) BglII (697) HindIII (722) stop codons BlpI (734) NheI (842) BmtI (846) PvuII (994) BspEI (1090) PasI (1326) MscI (1361) BtgI - NcoI - StyI (1395) XbaI (1698) PflFI - Tth111I (1858) BsaAI (1865) BstZ17I (1884) NdeI (1934) BstAPI (1935) pQE-16 3996 bp
BbsI  (3982)
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  (3980)
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  (3926)
1 site
G A C G T C C T G C A G
ZraI  (3924)
1 site
G A C G T C C T G C A G
FspI  (3226)
1 site
T G C G C A A C G C G T
NmeAIII  (3152)
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  (3124)
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  (3084)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
AhdI  (3004)
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.
PspFI  (2419)
1 site
C C C A G C G G G T C G
BseYI  (2415)
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.
PciI  (2111)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (1995)
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  (1995)
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.
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
BmeT110I  (2)
1 site
C Y C G R G G R G C Y 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
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.
SexAI  (301)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
Eco53kI  (391)
1 site
G A G C T C C T C G A G
BanII  (393)
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  (393)
1 site
G A G C T C C T C G A G
BstXI  (411)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
AflII  (423)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
EcoNI  (634)
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.
BglII  (697)
1 site
A G A T C T T C T A G A
HindIII  (722)
1 site
A A G C T T T T C G A A
BlpI  (734)
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  (842)
1 site
G C T A G C C G A T C G
BmtI  (846)
1 site
G C T A G C C G A T C G
PvuII  (994)
1 site
C A G C T G G T C G A C
BspEI  (1090)
1 site
T C C G G A A G G C C T
PasI  (1326)
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.
MscI  (1361)
1 site
T G G C C A A C C G G T
BtgI  (1395)
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  (1395)
1 site
C C A T G G G G T A C C
StyI  (1395)
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  (1698)
1 site
T C T A G A A G A T C T
PflFI  (1858)
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  (1858)
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  (1865)
1 site
Y A C G T R R T G C A Y
BstZ17I  (1884)
1 site
G T A T A C C A T A T G
NdeI  (1934)
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  (1935)
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.
AmpR
2931 .. 3791  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2931 .. 3722  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2931 .. 3791  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3723 .. 3791  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2931 .. 3791  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
881 .. 1540  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
881 .. 1540  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2172 .. 2760  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2172 .. 2760  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
DHFR
133 .. 690  =  558 bp
186 amino acids  =  21.5 kDa
Product: mouse dihydrofolate reductase
DHFR
133 .. 690  =  558 bp
186 amino acids  =  21.5 kDa
Product: mouse dihydrofolate reductase
AmpR promoter
3792 .. 3896  =  105 bp
AmpR promoter
3792 .. 3896  =  105 bp
lambda t0 terminator
743 .. 837  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
743 .. 837  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1605 .. 1691  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
1605 .. 1691  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
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
703 .. 720  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
703 .. 720  =  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
727 .. 737  =  11 bp
stop codons in all three reading frames
stop codons
727 .. 737  =  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
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).
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