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pTriEx™-4 Hygro

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

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pTriEx-4 Hygro Sequence and MappTriEx-4 Hygro.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 AanI (5197) AanI (4640) AanI (320) AanI (409) 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) PstI - SbfI (2325) BstBI (2342) NotI (2352) PvuII (2362) BstZ17I (2367) PaeR7I - XhoI (2414) Bsu36I (2456) pTriEx™-4 Hygro 6888 bp
AanI  (5197)
4 sites
T T A T A A A A T A T T
AanI  (4640)
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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.
PstI  (2325)
1 site
C T G C A G G A C G T C
SbfI  (2325)
1 site
C C T G C A G G G G A C G T C C
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 2
1863 .. 4272  =  2410 bp
Exon 2
1863 .. 4272  =  2410 bp
AmpR
5966 .. 6826  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5966 .. 6757  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5966 .. 6826  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6758 .. 6826  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5966 .. 6826  =  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)
4451 .. 5156  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4451 .. 5156  =  706 bp
contains part of ORF1629
ori
5366 .. 5953  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5366 .. 5953  =  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
intron
1699 .. 1862  =  164 bp
intron
1699 .. 1862  =  164 bp
Exon 1
1597 .. 1698  =  102 bp
Exon 1
1597 .. 1698  =  102 bp
T7 terminator
4391 .. 4438  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
4391 .. 4438  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
HygR
2999 .. 4018  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
2999 .. 4018  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
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
4568 .. 5156  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4568 .. 5156  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
IRES
2497 .. 2960  =  464 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
2497 .. 2960  =  464 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
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
MCS
2252 .. 2370  =  119 bp
multiple cloning site
MCS
2252 .. 2370  =  119 bp
multiple cloning site
rabbit globin terminator
4270 .. 4381  =  112 bp
rabbit globin terminator
4270 .. 4381  =  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
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
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
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).
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
T7 promoter
1931 .. 1949  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1931 .. 1949  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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
thrombin site
2207 .. 2224  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
2207 .. 2224  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
RBS
2101 .. 2106  =  6 bp
ribosome binding site
RBS
2101 .. 2106  =  6 bp
ribosome binding site
poly(A) signal
4248 .. 4253  =  6 bp
poly(A) signal
4248 .. 4253  =  6 bp
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
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
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