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pIRESneo3

Mammalian IRES-containing vector with a neomycin (G418) resistance marker for expressing two genes from the same bicistronic transcript.

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pIRESneo3.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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SgrDI (5273) SspI (5156) PvuI (4722) BpmI (4422) AhdI (4352) BstZ17I (3080) PsiI (2914) BclI * (2788) XbaI (2778) Bpu10I (180) NruI (208) SpeI (249) NdeI (484) SnaBI (590) BspDI * - ClaI * (910) EcoRV (914) NheI (927) BmtI (931) AfeI (932) AflII (934) StuI (940) AgeI (948) BsiWI (954) BspEI (962) BsmBI (964) BstBI (969) EcoRI (971) BamHI (977) NotI (984) BstXI (1011) AleI (1242) NsiI (1318) PspOMI (1466) ApaI (1470) AvrII (1504) PmlI (1669) AarI (1692) DraIII (1716) BmgBI (1896) AvaI - BsoBI - TspMI - XmaI (1938) BmeT110I (1939) SmaI (1940) KasI (2098) NarI (2099) SfoI (2100) PluTI (2102) PflFI - Tth111I (2217) BssHII (2496) NgoMIV (2599) NaeI (2601) RsrII (2615) pIRESneo3 5275 bp
SgrDI  (5273)
1 site
C G T C G A C G G C A G C T G C
SspI  (5156)
1 site
A A T A T T T T A T A A
PvuI  (4722)
1 site
C G A T C G G C T A G C
BpmI  (4422)
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.
AhdI  (4352)
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.
BstZ17I  (3080)
1 site
G T A T A C C A T A T G
PsiI  (2914)
1 site
T T A T A A A A T A T T
BclI  (2788)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
XbaI  (2778)
1 site
T C T A G A A G A T C T
Bpu10I  (180)
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.
NruI  (208)
1 site
T C G C G A A G C G C T
SpeI  (249)
1 site
A C T A G T T G A T C A
NdeI  (484)
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  (590)
1 site
T A C G T A A T G C A T
BspDI  (910)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (910)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoRV  (914)
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.
NheI  (927)
1 site
G C T A G C C G A T C G
BmtI  (931)
1 site
G C T A G C C G A T C G
AfeI  (932)
1 site
A G C G C T T C G C G A
AflII  (934)
1 site
C T T A A G G A A T T C
StuI  (940)
1 site
A G G C C T T C C G G A
AgeI  (948)
1 site
A C C G G T T G G C C A
BsiWI  (954)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BspEI  (962)
1 site
T C C G G A A G G C C T
BsmBI  (964)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BstBI  (969)
1 site
T T C G A A A A G C T T
EcoRI  (971)
1 site
G A A T T C C T T A A G
BamHI  (977)
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.
NotI  (984)
1 site
G C G G C C G C C G C C G G C G
BstXI  (1011)
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.
AleI  (1242)
1 site
C A C N N N N G T G G T G N N N N C A C
NsiI  (1318)
1 site
A T G C A T T A C G T A
PspOMI  (1466)
1 site
G G G C C C C C C G G G
ApaI  (1470)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (1504)
1 site
C C T A G G G G A T C C
PmlI  (1669)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (1692)
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.
DraIII  (1716)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BmgBI  (1896)
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.
AvaI  (1938)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1938)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (1938)
1 site
C C C G G G G G G C C C
XmaI  (1938)
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.
BmeT110I  (1939)
1 site
C Y C G R G G R G C Y C
SmaI  (1940)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KasI  (2098)
1 site
G G C G C C C C G C G G
NarI  (2099)
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  (2100)
1 site
G G C G C C C C G C G G
PluTI  (2102)
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.
PflFI  (2217)
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  (2217)
1 site
G A C N N N G T C C T G N N N C A G