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Plasmid Files

pIRES

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

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pIRES Sequence and MappIRES.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BglII (6100) AlwNI (5679) AhdI (5200) BpmI (5131) PfoI (4161) BstXI (3945) BstBI (3906) RsrII (3740) BssHII (3621) PflFI - Tth111I (3342) BbeI (3227) SfoI (3225) NarI (3224) KasI (3223) BsrGI (96) SpeI (152) CMV enhancer NdeI (387) SnaBI (493) AsiSI (664) Eco53kI (727) SacI (729) NheI (1085) BmtI (1089) PaeR7I - XhoI (1091) MCS A EcoRI (1096) MluI (1102) PspOMI (1265) ApaI (1269) PmlI (1468) AarI (1491) PflMI (1605) PciI (1642) BmgBI (1695) SmaI (1739) XbaI (1747) SalI (1753) AccI (1754) SmaI (1760) NotI (1764) T3 promoter AanI (1910) HpaI (1930) MfeI (1939) AanI (2474) SexAI * (2797) SfiI (2983) StuI (3029) pIRES 6105 bp
BglII  (6100)
1 site
A G A T C T T C T A G A
AlwNI  (5679)
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  (5200)
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  (5131)
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.
PfoI  (4161)
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.
BstXI  (3945)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BstBI  (3906)
1 site
T T C G A A A A G C T T
RsrII  (3740)
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  (3621)
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  (3342)
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  (3342)
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.
BbeI  (3227)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the BbeI
recognition sequence.
SfoI  (3225)
1 site
G G C G C C C C G C G G
NarI  (3224)
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.
KasI  (3223)
1 site
G G C G C C C C G C G G
BsrGI  (96)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
SpeI  (152)
1 site
A C T A G T T G A T C A
NdeI  (387)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SnaBI  (493)
1 site
T A C G T A A T G C A T
AsiSI  (664)
1 site
G C G A T C G C C G C T A G C G
Eco53kI  (727)
1 site
G A G C T C C T C G A G
SacI  (729)
1 site
G A G C T C C T C G A G
NheI  (1085)
1 site
G C T A G C C G A T C G
BmtI  (1089)
1 site
G C T A G C C G A T C G
PaeR7I  (1091)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1091)
1 site
C T C G A G G A G C T C
EcoRI  (1096)
1 site
G A A T T C C T T A A G
MluI  (1102)
1 site
A C G C G T T G C G C A
PspOMI  (1265)
1 site
G G G C C C C C C G G G
ApaI  (1269)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PmlI  (1468)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (1491)
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  (1605)
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.
PciI  (1642)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BmgBI  (1695)
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.
SmaI  (1739)
2 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
XbaI  (1747)
1 site
T C T A G A A G A T C T
SalI  (1753)
1 site
G T C G A C C A G C T G
AccI  (1754)
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.
SmaI  (1760)
2 sites
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NotI  (1764)
1 site
G C G G C C G C C G C C G G C G
AanI  (1910)
2 sites
T T A T A A A A T A T T
HpaI  (1930)
1 site
G T T A A C C A A T T G
MfeI  (1939)
1 site
C A A T T G G T T A A C
AanI  (2474)
2 sites
T T A T A A A A T A T T
SexAI  (2797)
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.
SfiI  (2983)
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.
StuI  (3029)
1 site
A G G C C T T C C G G A
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4413 .. 4481  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4482 .. 5273  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4413 .. 5273  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
3096 .. 3890  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
3096 .. 3890  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
5444 .. 6032  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5444 .. 6032  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
1152 .. 1725  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
1152 .. 1725  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
f1 ori
2116 .. 2571  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2116 .. 2571  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2688 .. 3045  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2688 .. 3045  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
213 .. 517  =  305 bp
human cytomegalovirus immediate early enhancer
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
890 .. 1022  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
1809 .. 1930  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1809 .. 1930  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4308 .. 4412  =  105 bp
AmpR promoter
4308 .. 4412  =  105 bp
poly(A) signal
3954 .. 4002  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3954 .. 4002  =  49 bp
synthetic polyadenylation signal
minimal CMV promoter
691 .. 729  =  39 bp
human cytomegalovirus (CMV) immediate early
promoter
minimal CMV promoter
691 .. 729  =  39 bp
human cytomegalovirus (CMV) immediate early
promoter
MCS B
1737 .. 1770  =  34 bp
multiple cloning site
MCS B
1737 .. 1770  =  34 bp
multiple cloning site
MCS A
1085 .. 1107  =  23 bp
multiple cloning site
MCS A
1085 .. 1107  =  23 bp
multiple cloning site
T3 promoter
1774 .. 1791  =  18 bp
promoter for bacteriophage T3 RNA polymerase
(shorter by one base than the standard T3
promoter)
T3 promoter
1774 .. 1791  =  18 bp
promoter for bacteriophage T3 RNA polymerase
(shorter by one base than the standard T3
promoter)
SV40 ori
2896 .. 3031  =  136 bp
SV40 origin of replication
SV40 ori
2896 .. 3031  =  136 bp
SV40 origin of replication
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1067 .. 1085  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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