pTriEx-2 Neo

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

Sequence Author: MilliporeSigma (Novagen)

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Explore Over 2.7k Plasmids: Insect Cell Vectors | More Plasmid Sets
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AanI (5129) AanI (4572) AanI (320) AanI (409) NcoI (2331) XcmI (2357) TspMI - XmaI (2433) SmaI (2435) PshAI (2475) BseRI (2479) EcoRV (2499) Eco53kI (2506) SacI (2508) BamHI (2512) EcoRI (2518) BglII (2526) AscI (2534) SbfI (2544) AgeI (2555) BstBI (2561) NotI (2571) PvuII (2581) BstZ17I (2586) PaeR7I - XhoI (2633) Bsu36I (2675) pTriEx™-2 Neo 6820 bp
AanI  (5129)
4 sites
T T A T A A A A T A T T
AanI  (4572)
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  (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.
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
SbfI  (2544)
1 site
C C T G C A G G G G A C G T C C
AgeI  (2555)
1 site
A C C G G T T G G C C A
BstBI  (2561)
1 site
T T C G A A A A G C T T
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
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
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.
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 .. 4204  =  2123 bp
Exon 2
2082 .. 4204  =  2123 bp
AmpR
5898 .. 6758  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5898 .. 6689  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5898 .. 6758  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6690 .. 6758  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5898 .. 6758  =  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)
4383 .. 5088  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4383 .. 5088  =  706 bp
contains part of ORF1629
ori
5298 .. 5885  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5298 .. 5885  =  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
4323 .. 4370  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4323 .. 4370  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
NeoR/KanR
3259 .. 4062  =  804 bp
267 amino acids  =  29.2 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3259 .. 4062  =  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
4500 .. 5088  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4500 .. 5088  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
IRES
2761 .. 3223  =  463 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2761 .. 3223  =  463 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
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
4202 .. 4313  =  112 bp
rabbit globin terminator
4202 .. 4313  =  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
4180 .. 4185  =  6 bp
poly(A) signal
4180 .. 4185  =  6 bp
MCS
2471 .. 2589  =  119 bp
multiple cloning site
MCS
2471 .. 2589  =  119 bp
multiple cloning site
ORF:  1315 .. 1704  =  390 bp
ORF:  129 amino acids  =  13.9 kDa
ORF:  3133 .. 4062  =  930 bp
ORF:  309 amino acids  =  33.9 kDa
ORF:  6028 .. 6294  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6808 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 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:  3440 .. 3826  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  3577 .. 3831  =  255 bp
ORF:  84 amino acids  =  9.7 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:  4500 .. 5117  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  5898 .. 6758  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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