pTriEx-2

Insect, bacterial, and mammalian vector for expressing proteins with a cleavable N-terminal 6xHis-S-Tag cassette and a C-terminal HSV-8xHis cassette.

Sequence Author: MilliporeSigma (Novagen)

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AanI (3766) baculovirus recombination region (ORF1629) AanI (3209) Bsu36I (2675) DraIII (2665) PaeR7I - XhoI (2633) AanI (320) AanI (409) baculovirus recombination region (lef2/ORF603) NcoI (2331) XcmI (2357) PflMI (2401) TspMI - XmaI (2433) SmaI (2435) PshAI (2475) BseRI (2479) EcoRV (2499) Eco53kI (2506) SacI (2508) BamHI (2512) EcoRI (2518) BglII (2526) AscI (2534) PstI - SbfI (2544) Acc65I (2552) AgeI (2555) KpnI (2556) BstBI (2561) HindIII (2564) NotI (2571) PvuII (2581) BstZ17I (2586) PmlI (2591) pTriEx™-2 5457 bp
AanI  (3766)
4 sites
T T A T A A A A T A T T
AanI  (3209)
4 sites
T T A T A A A A T A T T
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.
DraIII  (2665)
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.
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
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.
PflMI  (2401)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI 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
PstI  (2544)
1 site
C T G C A G G A C G T C
SbfI  (2544)
1 site
C C T G C A G G G G A C G T C C
Acc65I  (2552)
1 site
G G T A C C C C A T G G
AgeI  (2555)
1 site
A C C G G T T G G C C A
KpnI  (2556)
1 site
G G T A C C C C A T G G
BstBI  (2561)
1 site
T T C G A A A A G C T T
HindIII  (2564)
1 site
A A G C T T T T C G A A
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
PmlI  (2591)
1 site
C A C G T G G T G C A 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 .. 2841  =  760 bp
Exon 2
2082 .. 2841  =  760 bp
AmpR
4535 .. 5395  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4535 .. 5326  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4535 .. 5395  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5327 .. 5395  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4535 .. 5395  =  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)
3020 .. 3725  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3020 .. 3725  =  706 bp
contains part of ORF1629
ori
3935 .. 4522  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3935 .. 4522  =  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
2960 .. 3007  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2960 .. 3007  =  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:  2333 .. 2665  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  4665 .. 4931  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5445 .. 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:  2492 .. 2803  =  312 bp
ORF:  103 amino acids  =  11.8 kDa
ORF:  3137 .. 3754  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  4535 .. 5395  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

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