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Plasmid Files

pQE-T7-1

Vector for expressing N-terminally 10xHis-tagged proteins in bacteria using the QIAgenes system.

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pQE-T7-1 Sequence and MappQE-T7-1.dna
Map and Sequence File   
Sequence Author:  Qiagen
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 DraIII (5075) PsiI (4947) AsiSI - PvuI (4375) SmaI (4249) TspMI - XmaI (4247) BspDI - ClaI (4066) NruI (4032) AcuI (3721) AlwNI (3589) BssS α I (3346) PciI (3173) BspQI - SapI (3057) TatI (2980) BstZ17I (2947) AccI (2946) PflFI - Tth111I (2921) StyI (57) BlpI (80) 6xHis PaeR7I - PspXI - XhoI (158) PstI (178) BmgBI (181) Acc65I (185) KpnI (189) EagI - NotI - SacII (193) Eco53kI (204) SacI (206) NdeI (213) ATG RBS XbaI (287) lac operator T7 promoter BglII (353) SgrAI (394) SphI (550) BstAPI (758) MluI (1075) BclI * (1089) BstEII (1256) NmeAIII (1281) PspOMI (1282) ApaI (1286) BssHII (1486) EcoRV (1525) HincII - HpaI (1581) PshAI (1920) BglI (2139) FspI - FspAI (2157) PpuMI (2182) pQE-T7-1 5317 bp
DraIII  (5075)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
PsiI  (4947)
1 site
T T A T A A A A T A T T
AsiSI  (4375)
1 site
G C G A T C G C C G C T A G C G
PvuI  (4375)
1 site
C G A T C G G C T A G C
SmaI  (4249)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (4247)
1 site
C C C G G G G G G C C C
XmaI  (4247)
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.
BspDI  (4066)
1 site
A T C G A T T A G C T A
ClaI  (4066)
1 site
A T C G A T T A G C T A
NruI  (4032)
1 site
T C G C G A A G C G C T
AcuI  (3721)
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  (3589)
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.
BssSαI  (3346)
1 site
C A C G A G G T G C T C
PciI  (3173)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3057)
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  (3057)
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.
TatI  (2980)
1 site
W G T A C W W C A T G W
BstZ17I  (2947)
1 site
G T A T A C C A T A T G
AccI  (2946)
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.
PflFI  (2921)
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  (2921)
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.
StyI  (57)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BlpI  (80)
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.
PaeR7I  (158)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site
V C T C G A G B B G A G C T C V
XhoI  (158)
1 site
C T C G A G G A G C T C
PstI  (178)
1 site
C T G C A G G A C G T C
BmgBI  (181)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
Acc65I  (185)
1 site
G G T A C C C C A T G G
KpnI  (189)
1 site
G G T A C C C C A T G G
EagI  (193)
1 site
C G G C C G G C C G G C
NotI  (193)
1 site
G C G G C C G C C G C C G G C G
SacII  (193)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
Eco53kI  (204)
1 site
G A G C T C C T C G A G
SacI  (206)
1 site
G A G C T C C T C G A G
NdeI  (213)
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.
XbaI  (287)
1 site
T C T A G A A G A T C T
BglII  (353)
1 site
A G A T C T T C T A G A
SgrAI  (394)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
SphI  (550)
1 site
G C A T G C C G T A C G
BstAPI  (758)
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.
MluI  (1075)
1 site
A C G C G T T G C G C A
BclI  (1089)
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.
BstEII  (1256)
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.
NmeAIII  (1281)
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).
PspOMI  (1282)
1 site
G G G C C C C C C G G G
ApaI  (1286)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BssHII  (1486)
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.
EcoRV  (1525)
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.
HincII  (1581)
1 site
G T Y R A C C A R Y T G
HpaI  (1581)
1 site
G T T A A C C A A T T G
PshAI  (1920)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BglI  (2139)
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.
FspI  (2157)
1 site
T G C G C A A C G C G T
FspAI  (2157)
1 site
R T G C G C A Y Y A C G C G T R
PpuMI  (2182)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
lacI
725 .. 1807  =  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
725 .. 1807  =  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
3944 .. 4759  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
KanR
3944 .. 4759  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin
ori
3234 .. 3822  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3234 .. 3822  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
4851 .. 5306  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4851 .. 5306  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
2616 .. 2807  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
2616 .. 2807  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
lacI promoter
647 .. 724  =  78 bp
lacI promoter
647 .. 724  =  78 bp
MCS
158 .. 217  =  60 bp
multiple cloning site
MCS
158 .. 217  =  60 bp
multiple cloning site
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
lac operator
295 .. 319  =  25 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
295 .. 319  =  25 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).
T7 promoter
320 .. 338  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
320 .. 338  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
140 .. 157  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
RBS
258 .. 263  =  6 bp
ribosome binding site
RBS
258 .. 263  =  6 bp
ribosome binding site
ATG
248 .. 250  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
248 .. 250  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
10xHis
215 .. 244  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
10xHis
215 .. 244  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
stop codons
166 .. 171  =  6 bp
two tandem stop codons
stop codons
166 .. 171  =  6 bp
two tandem stop codons
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