pTriEx-3 Hygro

Insect, bacterial, and mammalian vector with a CMV promoter and a hygromycin resistance marker, for expressing proteins with a C-terminal HSV-8xHis cassette.

Sequence Author: MilliporeSigma (Novagen)

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AanI (5041) AanI (4484) AanI (320) AanI (409) CMV enhancer NcoI (2112) EcoRV (2121) TspMI - XmaI (2124) SmaI (2126) Eco53kI (2131) SacI (2133) BamHI (2137) EcoRI (2143) BglII (2151) AscI - BssHII (2159) PstI - SbfI (2169) BstBI (2186) NotI (2196) PvuII (2206) BstZ17I (2211) PaeR7I - XhoI (2258) Bsu36I (2300) pTriEx™-3 Hygro 6732 bp
AanI  (5041)
4 sites
T T A T A A A A T A T T
AanI  (4484)
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
EcoRV  (2121)
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.
TspMI  (2124)
1 site
C C C G G G G G G C C C
XmaI  (2124)
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  (2126)
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  (2131)
1 site
G A G C T C C T C G A G
SacI  (2133)
1 site
G A G C T C C T C G A G
BamHI  (2137)
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  (2143)
1 site
G A A T T C C T T A A G
BglII  (2151)
1 site
A G A T C T T C T A G A
AscI  (2159)
1 site
G G C G C G C C C C G C G C G G
BssHII  (2159)
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  (2169)
1 site
C T G C A G G A C G T C
SbfI  (2169)
1 site
C C T G C A G G G G A C G T C C
BstBI  (2186)
1 site
T T C G A A A A G C T T
NotI  (2196)
1 site
G C G G C C G C C G C C G G C G
PvuII  (2206)
1 site
C A G C T G G T C G A C
BstZ17I  (2211)
1 site
G T A T A C C A T A T G
PaeR7I  (2258)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2258)
1 site
C T C G A G G A G C T C
Bsu36I  (2300)
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 .. 4116  =  2254 bp
Exon 2
1863 .. 4116  =  2254 bp
AmpR
5810 .. 6670  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5810 .. 6601  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5810 .. 6670  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6602 .. 6670  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5810 .. 6670  =  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)
4295 .. 5000  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4295 .. 5000  =  706 bp
contains part of ORF1629
ori
5210 .. 5797  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5210 .. 5797  =  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
4235 .. 4282  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4235 .. 4282  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
HygR
2843 .. 3862  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
2843 .. 3862  =  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
4412 .. 5000  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4412 .. 5000  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
IRES
2341 .. 2804  =  464 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2341 .. 2804  =  464 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
HSV tag
2225 .. 2257  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2225 .. 2257  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
2264 .. 2287  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2264 .. 2287  =  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
4114 .. 4225  =  112 bp
rabbit globin terminator
4114 .. 4225  =  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
4092 .. 4097  =  6 bp
poly(A) signal
4092 .. 4097  =  6 bp
MCS
2112 .. 2219  =  108 bp
multiple cloning site
MCS
2112 .. 2219  =  108 bp
multiple cloning site
ORF:  2714 .. 3862  =  1149 bp
ORF:  382 amino acids  =  42.6 kDa
ORF:  5940 .. 6206  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6720 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  2287 .. 2715  =  429 bp
ORF:  142 amino acids  =  15.7 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:  2729 .. 3595  =  867 bp
ORF:  288 amino acids  =  32.8 kDa
ORF:  4412 .. 5029  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  5810 .. 6670  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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