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Plasmid Files

pQE-TriSystem Strep

Vector for expression in bacterial, insect, and vertebrate cells of proteins with a C-terminal Strep-Tag II tag.

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pQE-TriSystem Strep Sequence and MappQE-TriSystem Strep.dna
Map and Sequence File   
Sequence Author:  Qiagen
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 NcoI (1265) EcoRV (1274) TspMI - XmaI (1277) SmaI (1279) Eco53kI (1284) SacI (1286) BamHI (1290) EcoRI (1296) BglII (1304) BfuAI - BspMI (1311) AscI (1312) PstI - SbfI (1322) Acc65I (1330) KpnI (1334) HindIII (1342) EagI - NotI (1349) PvuII (1359) BstZ17I (1364) PmlI (1369) pQE-TriSystem Strep 5736 bp
NcoI  (1265)
1 site
C C A T G G G G T A C C
EcoRV  (1274)
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  (1277)
1 site
C C C G G G G G G C C C
XmaI  (1277)
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.
SmaI  (1279)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Eco53kI  (1284)
1 site
G A G C T C C T C G A G
SacI  (1286)
1 site
G A G C T C C T C G A G
BamHI  (1290)
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  (1296)
1 site
G A A T T C C T T A A G
BglII  (1304)
1 site
A G A T C T T C T A G A
BfuAI  (1311)
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  (1311)
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.
AscI  (1312)
1 site
G G C G C G C C C C G C G C G G
PstI  (1322)
1 site
C T G C A G G A C G T C
SbfI  (1322)
1 site
C C T G C A G G G G A C G T C C
Acc65I  (1330)
1 site
G G T A C C C C A T G G
KpnI  (1334)
1 site
G G T A C C C C A T G G
HindIII  (1342)
1 site
A A G C T T T T C G A A
EagI  (1349)
1 site
C G G C C G G C C G G C
NotI  (1349)
1 site
G C G G C C G C C G C C G G C G
PvuII  (1359)
1 site
C A G C T G G T C G A C
BstZ17I  (1364)
1 site
G T A T A C C A T A T G
PmlI  (1369)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
baculovirus recombination region (ORF1629)
1768 .. 3226  =  1459 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
1768 .. 3226  =  1459 bp
contains part of ORF1629
AmpR
3985 .. 4845  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3985 .. 4776  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3985 .. 4845  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4777 .. 4845  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3985 .. 4845  =  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)
4911 .. 2  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
4911 .. 2  =  828 bp
contains ORF603 and part of lef2
ori
3385 .. 3972  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3385 .. 3972  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
Exon 2
1005 .. 1589  =  585 bp
Exon 2
1005 .. 1589  =  585 bp
CMV enhancer
62 .. 367  =  306 bp
human cytomegalovirus (CMV) immediate early
enhancer
CMV enhancer
62 .. 367  =  306 bp
human cytomegalovirus (CMV) immediate early
enhancer
chicken β-actin promoter
373 .. 650  =  278 bp
chicken β-actin promoter
373 .. 650  =  278 bp
Intron
743 .. 1004  =  262 bp
Intron
743 .. 1004  =  262 bp
Exon 1
651 .. 742  =  92 bp
Exon 1
651 .. 742  =  92 bp
T7 terminator
1708 .. 1755  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1708 .. 1755  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
ORF1629
1885 .. 3226  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
1885 .. 3226  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
5073 .. 5678  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
5073 .. 5678  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
4911 .. 5035  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
4911 .. 5035  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
rabbit globin terminator
1587 .. 1698  =  112 bp
rabbit globin terminator
1587 .. 1698  =  112 bp
p10 promoter
1143 .. 1252  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
1143 .. 1252  =  110 bp
baculovirus promoter for expression in insect cells
MCS
1265 .. 1372  =  108 bp
multiple cloning site
MCS
1265 .. 1372  =  108 bp
multiple cloning site
T5 promoter
1073 .. 1116  =  44 bp
   Segment 1:  
   1073 .. 1086  =  14 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
   Segment 2:  -35  
   1087 .. 1092  =  6 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
   Segment 3:  
   1093 .. 1109  =  17 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
   Segment 4:  -10  
   1110 .. 1116  =  7 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
4 segments
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
Strep-Tag II
1381 .. 1404  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an
engineered form of streptavidin
Strep-Tag II
1381 .. 1404  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an
engineered form of streptavidin
stop codons
1411 .. 1421  =  11 bp
stop codons in all three reading frames
stop codons
1411 .. 1421  =  11 bp
stop codons in all three reading frames
RBS
1255 .. 1260  =  6 bp
ribosome binding site
RBS
1255 .. 1260  =  6 bp
ribosome binding site
poly(A) signal
1565 .. 1570  =  6 bp
poly(A) signal
1565 .. 1570  =  6 bp
lac operator
1092 .. 1108  =  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
1092 .. 1108  =  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).
ATG
1267 .. 1269  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1267 .. 1269  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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