pIRES

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

Sequence Author: Clontech (TaKaRa)

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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) PluTI (3227) SfoI (3225) NarI (3224) KasI (3223) BsrGI (96) SpeI (152) CMV enhancer NdeI (387) SnaBI (493) AsiSI (664) Eco53kI (727) SacI (729) I-PpoI (851) NheI (1085) BmtI (1089) PaeR7I - XhoI (1091) MCS A EcoRI (1096) MluI (1102) PspOMI (1265) ApaI (1269) PmlI (1468) PaqCI (1491) PflMI (1605) PciI (1642) BmgBI (1695) XbaI (1747) SalI (1753) AccI (1754) NotI (1764) T3 promoter HpaI (1930) MfeI (1939) CsiI - 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.
PluTI  (3227)
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  (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
I-PpoI  (851)
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  (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
PaqCI  (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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
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.
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.
NotI  (1764)
1 site
G C G G C C G C C G C C G G C G
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
CsiI  (2797)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
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