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Plasmid Files

pEF1α-IRES

IRES-containing vector for expressing two genes in mammalian cells from the same bicistronic transcript.

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pEF1alpha-IRES.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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AleI (130) AlwNI (6264) AhdI (5785) BpmI (5716) AseI (5610) AatII (4866) ZraI (4864) PfoI (4746) BstBI (4491) RsrII (4325) BssHII (4206) PflFI - Tth111I (3927) MscI (3891) SfiI (3568) SexAI * (3382) AgeI (231) FseI (861) Eco53kI (983) SacI (985) BbvCI - Bpu10I (1059) I-PpoI (1436) NheI (1670) BmtI (1674) EcoRI (1681) MluI (1687) PmlI (2053) AarI (2076) PflMI (2190) BmgBI (2280) XbaI (2332) SalI (2338) AccI (2339) NotI (2349) HpaI (2515) pEF1 α- IRES 6690 bp
AleI  (130)
1 site
C A C N N N N G T G G T G N N N N C A C
AlwNI  (6264)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (5785)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BpmI  (5716)
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.
AseI  (5610)
1 site
A T T A A T T A A T T A
AatII  (4866)
1 site
G A C G T C C T G C A G
ZraI  (4864)
1 site
G A C G T C C T G C A G
PfoI  (4746)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (4491)
1 site
T T C G A A A A G C T T
RsrII  (4325)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BssHII  (4206)
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.
PflFI  (3927)
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  (3927)
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.
MscI  (3891)
1 site
T G G C C A A C C G G T
SfiI  (3568)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
SexAI  (3382)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AgeI  (231)
1 site
A C C G G T T G G C C A
FseI  (861)
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.
Eco53kI  (983)
1 site
G A G C T C C T C G A G
SacI  (985)
1 site
G A G C T C C T C G A G
BbvCI  (1059)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1059)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
I-PpoI  (1436)
1 site
C T C T C T T A A G G T A G C G A G A G A A T T C C A T C G

I-PpoI is a homing endonuclease that can recognize a variety of similar recognition sequences.
NheI  (1670)
1 site
G C T A G C C G A T C G
BmtI  (1674)
1 site
G C T A G C C G A T C G
EcoRI  (1681)
1 site
G A A T T C C T T A A G
MluI  (1687)
1 site
A C G C G T T G C G C A
PmlI  (2053)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (2076)
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 AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PflMI  (2190)
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.
BmgBI  (2280)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
XbaI  (2332)
1 site
T C T A G A A G A T C T
SalI  (2338)
1 site
G T C G A C C A G C T G
AccI  (2339)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
NotI  (2349)
1 site
G C G G C C G C C G C C G G C G
HpaI  (2515)
1 site
G T T A A C C A A T T G
EF-1α promoter
154 .. 1335  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
154 .. 1335  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
Amp
4998 .. 5858  =  861 bp
286 amino acids  =  31.6 kDa
Amp
4998 .. 5858  =  861 bp
286 amino acids  =  31.6 kDa
Kan
3681 .. 4475  =  795 bp
264 amino acids  =  29.0 kDa
Kan
3681 .. 4475  =  795 bp
264 amino acids  =  29.0 kDa
ori
6029 .. 6617  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6029 .. 6617  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
1737 .. 2310  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
1737 .. 2310  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
f1 origin
2701 .. 3156  =  456 bp
f1 origin
2701 .. 3156  =  456 bp
SV40 promoter
3273 .. 3630  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
3273 .. 3630  =  358 bp
SV40 enhancer and early promoter
chimeric intron
1475 .. 1607  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
chimeric intron
1475 .. 1607  =  133 bp
chimera between introns from human β-globin and immunoglobulin heavy chain genes
SV40 poly(A) signal
2394 .. 2515  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2394 .. 2515  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4893 .. 4997  =  105 bp
AmpR promoter
4893 .. 4997  =  105 bp
MCS B
2332 .. 2355  =  24 bp
multiple cloning site
MCS B
2332 .. 2355  =  24 bp
multiple cloning site
MCS A
1670 .. 1692  =  23 bp
multiple cloning site
MCS A
1670 .. 1692  =  23 bp
multiple cloning site
EF-1α intron A
384 .. 1326  =  943 bp
intron upstream of the start codon
EF-1α intron A
384 .. 1326  =  943 bp
intron upstream of the start codon
SV40 origin
3481 .. 3616  =  136 bp
SV40 origin
3481 .. 3616  =  136 bp
T7 promoter
1652 .. 1670  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1652 .. 1670  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  3853 .. 4239  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  713 .. 1150  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  792 .. 1025  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  3681 .. 4475  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  4998 .. 5858  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3990 .. 4526  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  428 .. 1075  =  648 bp
ORF:  215 amino acids  =  23.7 kDa
ORF:  5462 .. 5728  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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