pTriEx-1.1

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

Sequence Author: MilliporeSigma (Novagen)

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AanI (3610) baculovirus recombination region (ORF1629) AanI (3053) DraIII (2509) AanI (320) AanI (409) baculovirus recombination region (lef2/ORF603) NcoI (2331) EcoRV (2340) TspMI - XmaI (2343) SmaI (2345) Eco53kI (2350) SacI (2352) BamHI (2356) EcoRI (2362) BglII (2370) BfuAI - BspMI (2377) AscI (2378) PstI - SbfI (2388) Acc65I (2396) AgeI (2399) KpnI (2400) BstBI (2405) HindIII (2408) NotI (2415) PvuII (2425) BstZ17I (2430) PmlI (2435) PaeR7I - XhoI (2477) pTriExâ„¢-1.1 5301 bp
AanI  (3610)
4 sites
T T A T A A A A T A T T
AanI  (3053)
4 sites
T T A T A A A A T A T T
DraIII  (2509)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
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
BfuAI  (2377)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (2377)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
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
Acc65I  (2396)
1 site
G G T A C C C C A T G G
AgeI  (2399)
1 site
A C C G G T T G G C C A
KpnI  (2400)
1 site
G G T A C C C C A T G G
BstBI  (2405)
1 site
T T C G A A A A G C T T
HindIII  (2408)
1 site
A A G C T T T T C G A A
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
PmlI  (2435)
1 site
C A C G T G G T G C A C
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
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 .. 2685  =  604 bp
Exon 2
2082 .. 2685  =  604 bp
AmpR
4379 .. 5239  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4379 .. 5170  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4379 .. 5239  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5171 .. 5239  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4379 .. 5239  =  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)
2864 .. 3569  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
2864 .. 3569  =  706 bp
contains part of ORF1629
ori
3779 .. 4366  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3779 .. 4366  =  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
2804 .. 2851  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2804 .. 2851  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
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:  4509 .. 4775  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5289 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 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:  2300 .. 2647  =  348 bp
ORF:  115 amino acids  =  13.0 kDa
ORF:  2981 .. 3598  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  4379 .. 5239  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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