pQE-19

Bacterial vector for expressing DHFR fusion proteins with a C-terminal 6xHis tag. Similar to pQE-16 and pQE-22, but with a different reading frame after the DHFR.

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) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS ATG BamHI (121) CsiI - SexAI * (301) Eco53kI (391) BanII - SacI (393) BstXI (411) AflII (423) EcoNI (634) BglII (696) HindIII (721) stop codons BlpI (733) NheI (841) BmtI (845) PvuII (993) BspEI (1089) PasI (1325) MscI (1360) BtgI - NcoI - StyI (1394) XbaI (1698) PflFI - Tth111I (1858) BsaAI (1865) BstZ17I (1884) NdeI (1934) BstAPI (1935) pQE-19 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

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.
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
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.
CsiI  (301)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
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
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  (696)
1 site
A G A T C T T C T A G A
HindIII  (721)
1 site
A A G C T T T T C G A A
BlpI  (733)
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  (841)
1 site
G C T A G C C G A T C G
BmtI  (845)
1 site
G C T A G C C G A T C G
PvuII  (993)
1 site
C A G C T G G T C G A C
BspEI  (1089)
1 site
T C C G G A A G G C C T
PasI  (1325)
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  (1360)
1 site
T G G C C A A C C G G T
BtgI  (1394)
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  (1394)
1 site
C C A T G G G G T A C C
StyI  (1394)
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  =  31.5 kDa
2 segments
   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  =  31.5 kDa
2 segments
   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  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
880 .. 1539  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
880 .. 1539  =  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
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
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
742 .. 836  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
742 .. 836  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1604 .. 1690  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
1604 .. 1690  =  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
6xHis
702 .. 719  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
702 .. 719  =  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
726 .. 736  =  11 bp
stop codons in all three reading frames
stop codons
726 .. 736  =  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
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).
ORF:  115 .. 732  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  880 .. 1539  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3061 .. 3327  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  81 .. 314  =  234 bp
ORF:  77 amino acids  =  8.8 kDa
ORF:  2931 .. 3791  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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