pSF-CMV-RSV-FLuc AscI

Mammalian vector for co-expressing a gene together with firefly luciferase expressed from the Rous sarcoma virus promoter.

Sequence Author: Oxford Genetics

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PmeI (6324) rrnG terminator RsrII (6017) PflFI - Tth111I (5619) PmeI (5276) PpuMI - SanDI (5210) AscI (5119) BpmI (4765) SexAI * (4724) DraIII (4531) AgeI (4516) BlpI (4339) PluTI (4336) SfoI (4334) NarI (4333) KasI (4332) BsrGI (3875) ScaI (3550) PvuII (3459) BbsI (3443) XcmI (3381) SphI (3202) AsiSI (5) 5' β-globin insulator BglII (232) SnaBI (566) BglII (816) EagI - NotI (858) HindIII (869) Eco53kI (879) SacI (881) EcoRI (885) KpnI (901) NcoI (905) KpnI (915) EcoRV (921) AbsI - PaeR7I - PspXI - XhoI (928) XbaI (937) BseRI - BsgI (940) stop codons BspDI - ClaI (977) BamHI (986) StuI (996) NheI (1002) BmtI (1006) stop codons BtsI (1098) AanI - PsiI (1152) T7 terminator PstI - SbfI (1518) 3' β-globin insulator BstEII (1624) PacI (1650) SwaI (1776) AlwNI (2196) SwaI (2669) FseI (2775) AscI (2921) NruI (2978) MluI (2998) Bsu36I (3070) pSF-CMV-RSV-FLuc AscI 6443 bp
PmeI  (6324)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (6017)
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  (5619)
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  (5619)
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.
PmeI  (5276)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (5210)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
SanDI  (5210)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI sites may not be compatible.
AscI  (5119)
2 sites
G G C G C G C C C C G C G C G G
BpmI  (4765)
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.
SexAI  (4724)
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.
DraIII  (4531)
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.
AgeI  (4516)
1 site
A C C G G T T G G C C A
BlpI  (4339)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
PluTI  (4336)
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  (4334)
1 site
G G C G C C C C G C G G
NarI  (4333)
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  (4332)
1 site
G G C G C C C C G C G G
BsrGI  (3875)
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.
ScaI  (3550)
1 site
A G T A C T T C A T G A
PvuII  (3459)
1 site
C A G C T G G T C G A C
BbsI  (3443)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
XcmI  (3381)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SphI  (3202)
1 site
G C A T G C C G T A C G
AsiSI  (5)
1 site
G C G A T C G C C G C T A G C G
BglII  (232)
2 sites
A G A T C T T C T A G A
SnaBI  (566)
1 site
T A C G T A A T G C A T
BglII  (816)
2 sites
A G A T C T T C T A G A
EagI  (858)
1 site
C G G C C G G C C G G C
NotI  (858)
1 site
G C G G C C G C C G C C G G C G
HindIII  (869)
1 site
A A G C T T T T C G A A
Eco53kI  (879)
1 site
G A G C T C C T C G A G
SacI  (881)
1 site
G A G C T C C T C G A G
EcoRI  (885)
1 site
G A A T T C C T T A A G
KpnI  (901)
2 sites
G G T A C C C C A T G G
NcoI  (905)
1 site
C C A T G G G G T A C C
KpnI  (915)
2 sites
G G T A C C C C A T G G
EcoRV  (921)
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.
AbsI  (928)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (928)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (928)
1 site
V C T C G A G B B G A G C T C V
XhoI  (928)
1 site
C T C G A G G A G C T C
XbaI  (937)
1 site
T C T A G A A G A T C T
BseRI  (940)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BsgI  (940)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N )