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pIRES2 DsRed-Express2

IRES-containing bicistronic vector for expressing a gene together with the fluorescent protein DsRed-Express2.

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pIRES2 DsRed-Express2 Sequence and MappIRES2 DsRed-Express2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BsaI (4278) PfoI (4064) RsrII (3805) PflFI - Tth111I (3407) FspI (3391) PluTI (3292) SfoI (3290) NarI (3289) KasI (3288) BspDI * - ClaI * (3129) StuI (3110) SfiI (3064) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) EcoRI (629) SalI (639) AccI (640) SacII (652) TspMI - XmaI (656) SmaI (658) BamHI (660) AclI (690) XmnI (879) BmgBI (1210) BstXI (1253) AhdI (1433) SbfI (1594) AleI (1881) BssHII (1898) NotI (1933) XbaI * (1943) MfeI (2039) HpaI (2052) Bts α I (2128) AflII (2171) pIRES2 DsRed-Express2 5265 bp
BsaI  (4278)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
PfoI  (4064)
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.
RsrII  (3805)
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.
PflFI  (3407)
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  (3407)
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.
FspI  (3391)
1 site
T G C G C A A C G C G T
PluTI  (3292)
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.
SfoI  (3290)
1 site
G G C G C C C C G C G G
NarI  (3289)
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  (3288)
1 site
G G C G C C C C G C G G
BspDI  (3129)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3129)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (3110)
1 site
A G G C C T T C C G G A
SfiI  (3064)
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.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
BglII  (609)
1 site
A G A T C T T C T A G A
PaeR7I  (613)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (613)
1 site
C T C G A G G A G C T C
Eco53kI  (618)
1 site
G A G C T C C T C G A G
SacI  (620)
1 site
G A G C T C C T C G A G
EcoRI  (629)
1 site
G A A T T C C T T A A G
SalI  (639)
1 site
G T C G A C C A G C T G
AccI  (640)
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.
SacII  (652)
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.
TspMI  (656)
1 site
C C C G G G G G G C C C
XmaI  (656)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (658)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (660)
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.
AclI  (690)
1 site
A A C G T T T T G C A A
XmnI  (879)
1 site
G A A N N N N T T C C T T N N N N A A G
BmgBI  (1210)
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.
BstXI  (1253)
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.
AhdI  (1433)
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.
SbfI  (1594)
1 site
C C T G C A G G G G A C G T C C
AleI  (1881)
1 site
C A C N N N N G T G G T G N N N N C A C
BssHII  (1898)
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.
NotI  (1933)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1943)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (2039)
1 site
C A A T T G G T T A A C
HpaI  (2052)
1 site
G T T A A C C A A T T G
BtsαI  (2128)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
AflII  (2171)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
NeoR/KanR
3161 .. 3955  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
3161 .. 3955  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
DsRed-Express2
1254 .. 1931  =  678 bp
225 amino acids  =  25.7 kDa
Product: noncytotoxic tetrameric variant of DsRed
fluorescent protein
mammalian codon-optimized
DsRed-Express2
1254 .. 1931  =  678 bp
225 amino acids  =  25.7 kDa
Product: noncytotoxic tetrameric variant of DsRed
fluorescent protein
mammalian codon-optimized
ori
4563 .. 5151  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4563 .. 5151  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES2
667 .. 1253  =  587 bp
   Segment 1:  
   667 .. 1241  =  575 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
   Segment 2:  ATG  
   1242 .. 1244  =  3 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
   Segment 3:  
   1245 .. 1253  =  9 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
3 segments
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
f1 ori
2181 .. 2636  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2181 .. 2636  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2769 .. 3126  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2769 .. 3126  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
2053 .. 2174  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2053 .. 2174  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2663 .. 2767  =  105 bp
AmpR promoter
2663 .. 2767  =  105 bp
MCS
591 .. 665  =  75 bp
multiple cloning site
MCS
591 .. 665  =  75 bp
multiple cloning site
HSV TK poly(A) signal
4187 .. 4234  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
4187 .. 4234  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2977 .. 3112  =  136 bp
SV40 origin of replication
SV40 ori
2977 .. 3112  =  136 bp
SV40 origin of replication
ATG
1242 .. 1244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ATG
1242 .. 1244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
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