pTriEx-1.1 Hygro

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

Sequence Author: MilliporeSigma (Novagen)

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AanI (5260) AanI (4703) AanI (320) AanI (409) NcoI (2331) EcoRV (2340) TspMI - XmaI (2343) SmaI (2345) Eco53kI (2350) SacI (2352) BamHI (2356) EcoRI (2362) BglII (2370) AscI (2378) PstI - SbfI (2388) BstBI (2405) NotI (2415) PvuII (2425) BstZ17I (2430) PaeR7I - XhoI (2477) Bsu36I (2519) pTriEx™-1.1 Hygro 6951 bp
AanI  (5260)
4 sites
T T A T A A A A T A T T
AanI  (4703)
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
EcoRV  (2340)
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  (2343)
1 site
C C C G G G G G G C C C
XmaI  (2343)
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  (2345)
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  (2350)
1 site
G A G C T C C T C G A G
SacI  (2352)
1 site
G A G C T C C T C G A G
BamHI  (2356)
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  (2362)
1 site
G A A T T C C T T A A G
BglII  (2370)
1 site
A G A T C T T C T A G A
AscI  (2378)
1 site
G G C G C G C C C C G C G C G G
PstI  (2388)
1 site
C T G C A G G A C G T C
SbfI  (2388)
1 site
C C T G C A G G G G A C G T C C
BstBI  (2405)
1 site
T T C G A A A A G C T T
NotI  (2415)
1 site
G C G G C C G C C G C C G G C G
PvuII  (2425)
1 site
C A G C T G G T C G A C
BstZ17I  (2430)
1 site
G T A T A C C A T A T G
PaeR7I  (2477)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2477)
1 site
C T C G A G G A G C T C
Bsu36I  (2519)
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 .. 4335  =  2254 bp
Exon 2
2082 .. 4335  =  2254 bp
AmpR
6029 .. 6889  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6029 .. 6820  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6029 .. 6889  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6821 .. 6889  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6029 .. 6889  =  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)
4514 .. 5219  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4514 .. 5219  =  706 bp
contains part of ORF1629
ori
5429 .. 6016  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5429 .. 6016  =  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
4454 .. 4501  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4454 .. 4501  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
HygR
3062 .. 4081  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
3062 .. 4081  =  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
4631 .. 5219  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
4631 .. 5219  =  589 bp
195 amino acids  =  22.3 kDa
Product: baculovirus capsid-associated protein
required for viral replication
IRES
2560 .. 3023  =  464 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2560 .. 3023  =  464 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
HSV tag
2444 .. 2476  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2444 .. 2476  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
8xHis
2483 .. 2506  =  24 bp
8 amino acids  =  1.1 kDa
Product: 8xHis affinity tag
8xHis
2483 .. 2506  =  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
4333 .. 4444  =  112 bp
rabbit globin terminator
4333 .. 4444  =  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
4311 .. 4316  =  6 bp
poly(A) signal
4311 .. 4316  =  6 bp
MCS
2331 .. 2438  =  108 bp
multiple cloning site
MCS
2331 .. 2438  =  108 bp
multiple cloning site
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:  2933 .. 4081  =  1149 bp
ORF:  382 amino acids  =  42.6 kDa
ORF:  6159 .. 6425  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6939 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  2506 .. 2934  =  429 bp
ORF:  142 amino acids  =  15.7 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:  2948 .. 3814  =  867 bp
ORF:  288 amino acids  =  32.8 kDa
ORF:  4631 .. 5248  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  6029 .. 6889  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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