pTriEx-1.1 Neo

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

Sequence Author: MilliporeSigma (Novagen)

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ScaI (6295) BpmI (5885) AsiSI (4937) baculovirus recombination region (ORF1629) SwaI (4481) SphI (4225) BstAPI (4152) BspHI (3991) BspDI * - ClaI * (3955) PflFI - Tth111I (3358) MluI (399) BbvCI - Bpu10I (416) baculovirus recombination region (lef2/ORF603) FseI (1017) Exon 1 SacII (1965) XbaI (2034) PpuMI (2208) PacI (2280) NcoI (2331) EcoRV (2340) TspMI - XmaI (2343) SmaI (2345) Eco53kI (2350) SacI (2352) BamHI (2356) EcoRI (2362) BglII (2370) AscI (2378) SbfI (2388) AgeI (2399) BstBI (2405) NotI (2415) PvuII (2425) BstZ17I (2430) PaeR7I - XhoI (2477) Bsu36I (2519) AvrII (2645) PaqCI (2833) PflMI (2947) KasI (3239) NarI (3240) SfoI (3241) PluTI (3243) MscI (3322) pTriEx™-1.1 Neo 6664 bp
ScaI  (6295)
1 site
A G T A C T T C A T G A
BpmI  (5885)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AsiSI  (4937)
1 site
G C G A T C G C C G C T A G C G
SwaI  (4481)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
SphI  (4225)
1 site
G C A T G C C G T A C G
BstAPI  (4152)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BspHI  (3991)
1 site
T C A T G A A G T A C T
BspDI  (3955)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3955)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PflFI  (3358)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3358)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
MluI  (399)
1 site
A C G C G T T G C G C A
BbvCI  (416)
1 site
C C T C A G C G G A G T C G
Bpu10I  (416)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
FseI  (1017)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
SacII  (1965)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
XbaI  (2034)
1 site
T C T A G A A G A T C T
PpuMI  (2208)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
PacI  (2280)
1 site
T T A A T T A A A A T T A 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
SbfI  (2388)
1 site
C C T G C A G G G G A C G T C C
AgeI  (2399)
1 site
A C C G G T T G G C C A
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.
AvrII  (2645)
1 site
C C T A G G G G A T C C
PaqCI  (2833)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
PflMI  (2947)
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.
KasI  (3239)
1 site
G G C G C C C C G C G G
NarI  (3240)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (3241)
1 site
G G C G C C C C G C G G
PluTI  (3243)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
MscI  (3322)
1 site
T G G C C A A C C G G T
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 .. 4048  =  1967 bp
Exon 2
2082 .. 4048  =  1967 bp
AmpR
5742 .. 6602  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5742 .. 6533  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5742 .. 6602  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6534 .. 6602  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5742 .. 6602  =  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)
4227 .. 4932  =  706 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
4227 .. 4932  =  706 bp
contains part of ORF1629
ori
5142 .. 5729  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5142 .. 5729  =  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
4167 .. 4214  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
4167 .. 4214  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
ORF:  1315 .. 1704  =  390 bp
ORF:  129 amino acids  =  13.9 kDa
ORF:  2977 .. 3906  =  930 bp
ORF:  309 amino acids  =  33.9 kDa
ORF:  5872 .. 6138  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6652 .. 305  =  318 bp
ORF:  105 amino acids  =  12.1 kDa
ORF:  1547 .. 1798  =  252 bp
ORF:  83 amino acids  =  8.7 kDa
ORF:  3284 .. 3670  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  343 .. 948  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  3421 .. 3675  =  255 bp
ORF:  84 amino acids  =  9.7 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:  4344 .. 4961  =  618 bp
ORF:  205 amino acids  =  23.4 kDa
ORF:  5742 .. 6602  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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