pTriEx-4 Neo

Insect, bacterial, and mammalian vector with a CMV promoter and a neomycin resistance marker, encoding an N-terminal 6xHis-S-Tag and a C-terminal HSV-8xHis.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspTriEx-4 Neo
AanI (4910) AanI (4353) AanI (320) AanI (409) CMV enhancer NcoI (2112) XcmI (2138) TspMI - XmaI (2214) SmaI (2216) PshAI (2256) BseRI (2260) EcoRV (2280) Eco53kI (2287) SacI (2289) BamHI (2293) EcoRI (2299) BglII (2307) AscI - BssHII (2315) SbfI (2325) AgeI (2336) BstBI (2342) NotI (2352) PvuII (2362) BstZ17I (2367) PaeR7I - XhoI (2414) Bsu36I (2456) pTriEx™-4 Neo 6601 bp
AanI  (4910)
4 sites		 T T A T A A A A T A T T
AanI  (4353)
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
NcoI  (2112)
1 site		 C C A T G G G G T A C C
XcmI  (2138)
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.
TspMI  (2214)
1 site		 C C C G G G G G G C C C
XmaI  (2214)
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  (2216)
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  (2256)
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  (2260)
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  (2280)
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  (2287)
1 site		 G A G C T C C T C G A G
SacI  (2289)
1 site		 G A G C T C C T C G A G
BamHI  (2293)
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  (2299)
1 site		 G A A T T C C T T A A G
BglII  (2307)
1 site		 A G A T C T T C T A G A
AscI  (2315)
1 site		 G G C G C G C C C C G C G C G G
BssHII  (2315)
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.
SbfI  (2325)
1 site		 C C T G C A G G G G A C G T C C
AgeI  (2336)
1 site		 A C C G G T T G G C C A
BstBI  (2342)
1 site		 T T C G A A A A G C T T
NotI  (2352)
1 site		 G C G G C C G C C G C C G G C G
PvuII  (2362)
1 site		 C A G C T G G T C G A C
BstZ17I  (2367)
1 site		 G T A T A C C A T A T G
PaeR7I  (2414)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2414)
1 site		 C T C G A G G A G C T C
Bsu36I  (2456)
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.
Exon 1
1597 .. 1698  =  102 bp
Exon 1
1597 .. 1698  =  102 bp
intron
1699 .. 1862  =  164 bp
intron
1699 .. 1862  =  164 bp
Exon 2
1863 .. 3985  =  2123 bp
Exon 2
1863 .. 3985  =  2123 bp
AmpR
5679 .. 6539  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5679 .. 6470  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5679 .. 6539  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6471 .. 6539  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5679 .. 6539  =  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)
4164 .. 4869  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4164 .. 4869  =  706 bp
contains part of ORF1629
ori
5079 .. 5666  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5079 .. 5666  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
1076 .. 1379  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1076 .. 1379  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
1381 .. 1580  =  200 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
1381 .. 1580  =  200 bp
human cytomegalovirus (CMV) immediate early promoter
T7 terminator
4104 .. 4151  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4104 .. 4151  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
NeoR/KanR
3040 .. 3843  =  804 bp
267 amino acids  =  29.2 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3040 .. 3843  =  804 bp
267 amino acids  =  29.2 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
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
4281 .. 4869  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4281 .. 4869  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
IRES
2542 .. 3004  =  463 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2542 .. 3004  =  463 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
ATG
2114 .. 2116  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2114 .. 2116  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
2120 .. 2137  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2120 .. 2137  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
2147 .. 2191  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
2147 .. 2191  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
2207 .. 2224  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
2207 .. 2224  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
enterokinase site
2243 .. 2257  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
2243 .. 2257  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
HSV tag
2381 .. 2413  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2381 .. 2413  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
2420 .. 2443  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2420 .. 2443  =  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
3983 .. 4094  =  112 bp
rabbit globin terminator
3983 .. 4094  =  112 bp
p10 promoter
1990 .. 2099  =  110 bp
baculovirus promoter for expression in insect cells
p10 promoter
1990 .. 2099  =  110 bp
baculovirus promoter for expression in insect cells
lac operator
1950 .. 1974  =  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
1950 .. 1974  =  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
1931 .. 1949  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1931 .. 1949  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
2101 .. 2106  =  6 bp
ribosome binding site
RBS
2101 .. 2106  =  6 bp
ribosome binding site
poly(A) signal
3961 .. 3966  =  6 bp
poly(A) signal
3961 .. 3966  =  6 bp
MCS
2252 .. 2370  =  119 bp
multiple cloning site
MCS
2252 .. 2370  =  119 bp
multiple cloning site
ORF:  2914 .. 3843  =  930 bp
ORF:  309 amino acids  =  33.9 kDa
ORF:  5809 .. 6075  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6589 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  2114 .. 2446  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  3221 .. 3607  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  3358 .. 3612  =  255 bp
ORF:  84 amino acids  =  9.7 kDa
ORF:  387 .. 617  =  231 bp
ORF:  76 amino acids  =  9.2 kDa
ORF:  1587 .. 1841  =  255 bp
ORF:  84 amino acids  =  9.0 kDa
ORF:  4281 .. 4898  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  5679 .. 6539  =  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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