pTriEx-2

Insect, bacterial, and mammalian vector for expressing proteins with a cleavable N-terminal 6xHis-S-Tag cassette and a C-terminal HSV-8xHis cassette.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspTriEx-2
AanI (3766) AanI (3209) Bsu36I (2675) AanI (320) AanI (409) CMV enhancer NcoI (2331) XcmI (2357) PflMI (2401) TspMI - XmaI (2433) SmaI (2435) PshAI (2475) BseRI (2479) EcoRV (2499) Eco53kI (2506) SacI (2508) BamHI (2512) EcoRI (2518) BglII (2526) AscI (2534) PstI - SbfI (2544) Acc65I (2552) AgeI (2555) KpnI (2556) BstBI (2561) HindIII (2564) NotI (2571) PvuII (2581) BstZ17I (2586) PmlI (2591) PaeR7I - XhoI (2633) DraIII (2665) pTriEx™-2 5457 bp
AanI  (3766)
4 sites		 T T A T A A A A T A T T
AanI  (3209)
4 sites		 T T A T A A A A T A T T
Bsu36I  (2675)
1 site		 C C T N A G G G G A N T C C
Sticky ends from different Bsu36I sites may not be compatible.
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
NcoI  (2331)
1 site		 C C A T G G G G T A C C
XcmI  (2357)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PflMI  (2401)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
TspMI  (2433)
1 site		 C C C G G G G G G C C C
XmaI  (2433)
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  (2435)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
PshAI  (2475)
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.
BseRI  (2479)
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.
EcoRV  (2499)
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.
Eco53kI  (2506)
1 site		 G A G C T C C T C G A G
SacI  (2508)
1 site		 G A G C T C C T C G A G
BamHI  (2512)
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  (2518)
1 site		 G A A T T C C T T A A G
BglII  (2526)
1 site		 A G A T C T T C T A G A
AscI  (2534)
1 site		 G G C G C G C C C C G C G C G G
PstI  (2544)
1 site		 C T G C A G G A C G T C
SbfI  (2544)
1 site		 C C T G C A G G G G A C G T C C
Acc65I  (2552)
1 site		 G G T A C C C C A T G G
AgeI  (2555)
1 site		 A C C G G T T G G C C A
KpnI  (2556)
1 site		 G G T A C C C C A T G G
BstBI  (2561)
1 site		 T T C G A A A A G C T T
HindIII  (2564)
1 site		 A A G C T T T T C G A A
NotI  (2571)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (2581)
1 site		 C A G C T G G T C G A C
BstZ17I  (2586)
1 site		 G T A T A C C A T A T G
PmlI  (2591)
1 site		 C A C G T G G T G C A C
PaeR7I  (2633)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2633)
1 site		 C T C G A G G A G C T C
DraIII  (2665)
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.
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 .. 2841  =  760 bp
Exon 2
2082 .. 2841  =  760 bp
AmpR
4535 .. 5395  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4535 .. 5326  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4535 .. 5395  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5327 .. 5395  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4535 .. 5395  =  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)
3020 .. 3725  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3020 .. 3725  =  706 bp
contains part of ORF1629
ori
3935 .. 4522  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3935 .. 4522  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
1140 .. 1443  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1140 .. 1443  =  304 bp
human cytomegalovirus immediate early enhancer
chicken β-actin promoter
1449 .. 1726  =  278 bp
chicken β-actin promoter
1449 .. 1726  =  278 bp
T7 terminator
2960 .. 3007  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2960 .. 3007  =  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
3137 .. 3725  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
3137 .. 3725  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
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
6xHis
2339 .. 2356  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2339 .. 2356  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
2366 .. 2410  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
2366 .. 2410  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
2426 .. 2443  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
2426 .. 2443  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
enterokinase site
2462 .. 2476  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
2462 .. 2476  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
HSV tag
2600 .. 2632  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2600 .. 2632  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
2639 .. 2662  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2639 .. 2662  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
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
2839 .. 2950  =  112 bp
rabbit globin terminator
2839 .. 2950  =  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
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).
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2150 .. 2168  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
2320 .. 2325  =  6 bp
ribosome binding site
RBS
2320 .. 2325  =  6 bp
ribosome binding site
poly(A) signal
2817 .. 2822  =  6 bp
poly(A) signal
2817 .. 2822  =  6 bp
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:  2333 .. 2665  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  4665 .. 4931  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5445 .. 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:  2492 .. 2803  =  312 bp
ORF:  103 amino acids  =  11.8 kDa
ORF:  3137 .. 3754  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  4535 .. 5395  =  861 bp
ORF:  286 amino acids  =  31.6 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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