pQE-Trisystem-5

Vector for expressing N-terminally 10xHis-tagged proteins in insect and mammalian cells using the QIAgenes system.

Sequence Author: Qiagen

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HomePlasmidsQiagen VectorspQE-Trisystem-5
AanI (3601) AanI (3044) AanI (320) AanI (409) BspQI - SapI (2368) EcoRV (2382) TspMI - XmaI (2385) SmaI (2387) BamHI (2392) EcoRI (2398) BglII (2406) AscI (2414) PstI - SbfI (2424) BstBI (2429) HindIII (2432) PaeR7I - XhoI (2441) pQE-Trisystem-5 5292 bp
AanI  (3601)
4 sites		 T T A T A A A A T A T T
AanI  (3044)
4 sites		 T T A T A A A A T A T T
AanI  (320)
4 sites		 T T A T A A A A T A T T
AanI  (409)
4 sites		 T T A T A A A A T A T T
BspQI  (2368)
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  (2368)
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.
EcoRV  (2382)
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.
TspMI  (2385)
1 site		 C C C G G G G G G C C C
XmaI  (2385)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2387)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
BamHI  (2392)
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.
EcoRI  (2398)
1 site		 G A A T T C C T T A A G
BglII  (2406)
1 site		 A G A T C T T C T A G A
AscI  (2414)
1 site		 G G C G C G C C C C G C G C G G
PstI  (2424)
1 site		 C T G C A G G A C G T C
SbfI  (2424)
1 site		 C C T G C A G G G G A C G T C C
BstBI  (2429)
1 site		 T T C G A A A A G C T T
HindIII  (2432)
1 site		 A A G C T T T T C G A A
PaeR7I  (2441)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2441)
1 site		 C T C G A G G A G C T C
Exon 1
1727 .. 1819  =  93 bp
Exon 1
1727 .. 1819  =  93 bp
Intron
1820 .. 2081  =  262 bp
Intron
1820 .. 2081  =  262 bp
Exon 2
2082 .. 2676  =  595 bp
Exon 2
2082 .. 2676  =  595 bp
AmpR
4370 .. 5230  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4370 .. 5161  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4370 .. 5230  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5162 .. 5230  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4370 .. 5230  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
181 .. 1008  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (ORF1629)
2855 .. 3560  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
2855 .. 3560  =  706 bp
contains part of ORF1629
ori
3770 .. 4357  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3770 .. 4357  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
1138 .. 1443  =  306 bp
human cytomegalovirus (CMV) immediate early enhancer
CMV enhancer
1138 .. 1443  =  306 bp
human cytomegalovirus (CMV) immediate early enhancer
chicken β-actin promoter
1449 .. 1726  =  278 bp
chicken β-actin promoter
1449 .. 1726  =  278 bp
T7 terminator
2795 .. 2842  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2795 .. 2842  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
343 .. 948  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF1629
2972 .. 3560  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
2972 .. 3560  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
181 .. 305  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
rabbit globin terminator
2674 .. 2785  =  112 bp
rabbit globin terminator
2674 .. 2785  =  112 bp
p10 promoter
2209 .. 2318  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
2209 .. 2318  =  110 bp
baculovirus promoter for expression in insect cells
MCS
2368 .. 2446  =  79 bp
multiple cloning site
MCS
2368 .. 2446  =  79 bp
multiple cloning site
lac operator
2169 .. 2193  =  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
2169 .. 2193  =  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).
8xHis
2474 .. 2497  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2474 .. 2497  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
siRNA target site
2449 .. 2467  =  19 bp
siRNA target site
2449 .. 2467  =  19 bp
stop codons
2498 .. 2508  =  11 bp
stop codons in all three reading frames
stop codons
2498 .. 2508  =  11 bp
stop codons in all three reading frames
RBS
2321 .. 2326  =  6 bp
ribosome binding site
RBS
2321 .. 2326  =  6 bp
ribosome binding site
poly(A) signal
2652 .. 2657  =  6 bp
poly(A) signal
2652 .. 2657  =  6 bp
ATG
2333 .. 2335  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2333 .. 2335  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
10xHis
2339 .. 2368  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
can be processed with TAGZyme
10xHis
2339 .. 2368  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
can be processed with TAGZyme
ORF:  1315 .. 1704  =  390 bp
ORF:  129 amino acids  =  13.9 kDa
ORF:  1547 .. 1798  =  252 bp
ORF:  83 amino acids  =  8.7 kDa
ORF:  4500 .. 4766  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5280 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  387 .. 617  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  1671 .. 2060  =  390 bp
ORF:  129 amino acids  =  13.3 kDa
ORF:  2300 .. 2638  =  339 bp
ORF:  112 amino acids  =  12.7 kDa
ORF:  2972 .. 3589  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  4370 .. 5230  =  861 bp
ORF:  286 amino acids  =  31.6 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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