TAGZyme pQE-2

Vector with a lacIq repressor gene for bacterial expression of 6xHis-tagged proteins that can be processed by TAGZyme.

Sequence Author: Qiagen

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T5 promoter AvaI - BsoBI - PaeR7I - XhoI (1) AatII (4688) ZraI (4686) XmnI (4365) PvuI (4136) BglI (3886) BsaI (3827) AhdI (3766) AlwNI (3289) PciI (2873) BspQI - SapI (2757) BstZ17I (2646) PflFI - Tth111I (2620) PfoI (2517) FspAI (2453) PpuMI * (2436) PsiI (49) MfeI (59) lac operator EcoRI (88) RBS BseRI (115) ATG 6xHis NdeI (140) NsiI (146) SphI (148) Eco53kI (154) SacI (156) EagI - NotI (161) SacII (167) Acc65I (169) KpnI (173) PmlI (177) SalI (180) PstI (190) BfuAI - BspMI (193) HindIII (194) BlpI (206) NheI (314) BmtI (318) Bpu10I (339) BspEI (562) PasI (798) NcoI - StyI (867) KasI (1344) NarI * (1345) SfoI (1346) PluTI (1348) HpaI (1481) EcoRV (1537) BssHII (1572) PspOMI (1776) ApaI (1780) BstEII (1801) BclI * (1969) MluI (1983) TAGZyme pQE-2 4758 bp
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
AatII  (4688)
1 site		 G A C G T C C T G C A G
ZraI  (4686)
1 site		 G A C G T C C T G C A G
XmnI  (4365)
1 site		 G A A N N N N T T C C T T N N N N A A G
PvuI  (4136)
1 site		 C G A T C G G C T A G C
BglI  (3886)
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  (3827)
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  (3766)
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  (3289)
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  (2873)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (2757)
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  (2757)
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.
BstZ17I  (2646)
1 site		 G T A T A C C A T A T G
PflFI  (2620)
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  (2620)
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  (2517)
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  (2453)
1 site		 R T G C G C A Y Y A C G C G T R
PpuMI  (2436)
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.
NdeI  (140)
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.
NsiI  (146)
1 site		 A T G C A T T A C G T A
SphI  (148)
1 site		 G C A T G C C G T A C G
Eco53kI  (154)
1 site		 G A G C T C C T C G A G
SacI  (156)
1 site		 G A G C T C C T C G A G
EagI  (161)
1 site		 C G G C C G G C C G G C
NotI  (161)
1 site		 G C G G C C G C C G C C G G C G
SacII  (167)
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.
Acc65I  (169)
1 site		 G G T A C C C C A T G G
KpnI  (173)
1 site		 G G T A C C C C A T G G
PmlI  (177)
1 site		 C A C G T G G T G C A C
SalI  (180)
1 site		 G T C G A C C A G C T G
PstI  (190)
1 site		 C T G C A G G A C G T C
BfuAI  (193)
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  (193)
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  (194)
1 site		 A A G C T T T T C G A A
BlpI  (206)
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  (314)
1 site		 G C T A G C C G A T C G
BmtI  (318)
1 site		 G C T A G C C G A T C G
Bpu10I  (339)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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  (562)
1 site		 T C C G G A A G G C C T
PasI  (798)
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  (867)
1 site		 C C A T G G G G T A C C
StyI  (867)
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  (1344)
1 site		 G G C G C C C C G C G G
NarI  (1345)
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  (1346)
1 site		 G G C G C C C C G C G G
PluTI  (1348)
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  (1481)
1 site		 G T T A A C C A A T T G
EcoRV  (1537)
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  (1572)
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  (1776)
1 site		 G G G C C C C C C G G G
ApaI  (1780)
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  (1801)
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  (1969)
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  (1983)
1 site		 A C G C G T T G C G C A
lacI
1256 .. 2338  =  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
1256 .. 2338  =  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
3693 .. 4553  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3693 .. 4484  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3693 .. 4553  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4485 .. 4553  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3693 .. 4553  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
353 .. 1012  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
353 .. 1012  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2934 .. 3522  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2934 .. 3522  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
4554 .. 4658  =  105 bp
AmpR promoter
4554 .. 4658  =  105 bp
lambda t0 terminator
215 .. 309  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
215 .. 309  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1077 .. 1163  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
1077 .. 1163  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
lacIq promoter
2339 .. 2416  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
2339 .. 2416  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
MCS
139 .. 199  =  61 bp
multiple cloning site
MCS
139 .. 199  =  61 bp
multiple cloning site
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
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
6xHis
121 .. 138  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
121 .. 138  =  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
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
199 .. 209  =  11 bp
stop codons in all three reading frames
stop codons
199 .. 209  =  11 bp
stop codons in all three reading frames
ORF:  1222 .. 1473  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  3823 .. 4089  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  353 .. 1012  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  1236 .. 1499  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2088 .. 2405  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  3693 .. 4553  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1256 .. 2215  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
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Individual Sequences & Maps

pDrive
pQE-19
pQE-50
pQE-9
pDrive (linearized)
pQE-22
pQE-51
pQE-T7 (cloning region)
pIX 3.0
pQE-3
pQE-52
pQE-T7-1
pIX 4.0
pQE-30
pQE-6
pQE-T7-2
pQE-10
pQE-30 UA
pQE-60
pQE-TriSystem
pQE-100 DoubleTag
pQE-30 UA (linearized)
pQE-7
pQE-TriSystem His-Strep1
pQE-11
pQE-30 Xa
pQE-70
pQE-TriSystem His-Strep2
pQE-12
pQE-31
pQE-8
pQE-TriSystem Strep
pQE-13
pQE-32
pQE-80L
pQE-Trisystem-5
pQE-14
pQE-4
pQE-80L-Kan
pQE-Trisystem-6
pQE-15
pQE-40
pQE-81L
pQE-TriSystem-7
pQE-16
pQE-41
pQE-81L-Kan
pREP4
pQE-17
pQE-42
pQE-82L
TAGZyme pQE-1
pQE-18
pQE-5
pQE-82L-Kan
TAGZyme pQE-2

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