pQE-TriSystem His-Strep2

Vector for expression in bacterial, insect, and vertebrate cells of proteins with an N-terminal Strep-Tag II tag and a C-terminal 8xHis tag.

Sequence Author: Qiagen

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HomePlasmidsInsect Cell VectorspQE-TriSystem His-Strep2
AanI (5199) AanI (5110) AanI (3160) AanI (1111) NcoI (1265) TspMI - XmaI (1307) SmaI (1309) Eco53kI (1314) SacI (1316) BamHI (1320) EcoRI (1326) BglII (1334) BfuAI - BspMI (1341) AscI (1342) PstI - SbfI (1352) Acc65I (1360) KpnI (1364) HindIII (1372) EagI - NotI (1379) PvuII (1389) BstZ17I (1394) PmlI (1399) PaeR7I - XhoI (1441) AanI (2017) pQE-TriSystem His-Strep2 5796 bp
AanI  (5199)
5 sites		 T T A T A A A A T A T T
AanI  (5110)
5 sites		 T T A T A A A A T A T T
AanI  (3160)
5 sites		 T T A T A A A A T A T T
AanI  (1111)
5 sites		 T T A T A A A A T A T T
NcoI  (1265)
1 site		 C C A T G G G G T A C C
TspMI  (1307)
1 site		 C C C G G G G G G C C C
XmaI  (1307)
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  (1309)
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  (1314)
1 site		 G A G C T C C T C G A G
SacI  (1316)
1 site		 G A G C T C C T C G A G
BamHI  (1320)
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  (1326)
1 site		 G A A T T C C T T A A G
BglII  (1334)
1 site		 A G A T C T T C T A G A
BfuAI  (1341)
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  (1341)
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  (1342)
1 site		 G G C G C G C C C C G C G C G G
PstI  (1352)
1 site		 C T G C A G G A C G T C
SbfI  (1352)
1 site		 C C T G C A G G G G A C G T C C
Acc65I  (1360)
1 site		 G G T A C C C C A T G G
KpnI  (1364)
1 site		 G G T A C C C C A T G G
HindIII  (1372)
1 site		 A A G C T T T T C G A A
EagI  (1379)
1 site		 C G G C C G G C C G G C
NotI  (1379)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (1389)
1 site		 C A G C T G G T C G A C
BstZ17I  (1394)
1 site		 G T A T A C C A T A T G
PmlI  (1399)
1 site		 C A C G T G G T G C A C
PaeR7I  (1441)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1441)
1 site		 C T C G A G G A G C T C
AanI  (2017)
5 sites		 T T A T A A A A T A T T
baculovirus recombination region (ORF1629)
1828 .. 3286  =  1459 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
1828 .. 3286  =  1459 bp
contains part of ORF1629
Exon 1
651 .. 742  =  92 bp
Exon 1
651 .. 742  =  92 bp
Intron
743 .. 1004  =  262 bp
Intron
743 .. 1004  =  262 bp
Exon 2
1005 .. 1649  =  645 bp
Exon 2
1005 .. 1649  =  645 bp
AmpR
4045 .. 4905  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4045 .. 4836  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4045 .. 4905  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4837 .. 4905  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4045 .. 4905  =  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)
4971 .. 2  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
4971 .. 2  =  828 bp
contains ORF603 and part of lef2
ori
3445 .. 4032  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3445 .. 4032  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
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
T7 terminator
1768 .. 1815  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
1768 .. 1815  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ORF1629
1945 .. 3286  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
1945 .. 3286  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
5133 .. 5738  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
5133 .. 5738  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ATG
1267 .. 1269  =  3 bp
1 amino acid  =  149.2 Da
start codon
ATG
1267 .. 1269  =  3 bp
1 amino acid  =  149.2 Da
start codon
Strep-Tag II
1279 .. 1302  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
Strep-Tag II
1279 .. 1302  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin, an engineered form of streptavidin
HSV tag
1408 .. 1440  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
1408 .. 1440  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
1447 .. 1470  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
1447 .. 1470  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
lef2
4971 .. 5095  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
4971 .. 5095  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
rabbit globin terminator
1647 .. 1758  =  112 bp
rabbit globin terminator
1647 .. 1758  =  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
T5 promoter
1073 .. 1116  =  44 bp
4 segments
   Segment 1:  
   1073 .. 1086  =  14 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
4 segments
   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
4 segments
   Segment 3:  
   1093 .. 1109  =  17 bp
bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator
T5 promoter
1073 .. 1116  =  44 bp
4 segments
   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
stop codons
1471 .. 1481  =  11 bp
stop codons in all three reading frames
stop codons
1471 .. 1481  =  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
1625 .. 1630  =  6 bp
poly(A) signal
1625 .. 1630  =  6 bp
MCS
1307 .. 1446  =  140 bp
multiple cloning site
MCS
1307 .. 1446  =  140 bp
multiple cloning site
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).
ORF:  239 .. 628  =  390 bp
ORF:  129 amino acids  =  13.9 kDa
ORF:  4175 .. 4441  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  471 .. 725  =  255 bp
ORF:  84 amino acids  =  8.6 kDa
ORF:  1234 .. 1611  =  378 bp
ORF:  125 amino acids  =  14.2 kDa
ORF:  1945 .. 3198  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  4045 .. 4905  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  456 .. 983  =  528 bp
ORF:  175 amino acids  =  18.5 kDa
ORF:  5133 .. 5738  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  5177 .. 5407  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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