pSF-CMV-Ub-FLuc AscI

Mammalian vector for co-expressing a gene together with firefly luciferase expressed from the ubiquitin C promoter.

Sequence Author: Oxford Genetics

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PmeI (7081) rrnG terminator RsrII (6774) BsrDI (6491) PflFI - Tth111I (6376) PmeI (6033) AscI (5876) SexAI * (5481) AgeI (5273) BlpI (5096) FspI (4964) AleI (4640) BsrGI (4632) ScaI (4307) PvuII (4216) EcoNI (3869) AvrII (3865) AarI (3819) AanI (3792) AsiSI - PvuI (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 AanI (1152) MfeI (1181) T7 terminator PstI - SbfI (1518) 3' β-globin insulator BstEII (1624) PacI (1650) SwaI (1776) AlwNI (2196) SwaI (2669) FseI (2775) AscI (2921) SacII (2959) BsaI (3394) pSF-CMV-Ub-FLuc AscI 7200 bp
PmeI  (7081)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (6774)
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.
BsrDI  (6491)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (6376)
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  (6376)
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  (6033)
2 sites
G T T T A A A C C A A A T T T G
AscI  (5876)
2 sites
G G C G C G C C C C G C G C G G
SexAI  (5481)
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.
AgeI  (5273)
1 site
A C C G G T T G G C C A
BlpI  (5096)
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.
FspI  (4964)
1 site
T G C G C A A C G C G T
AleI  (4640)
1 site
C A C N N N N G T G G T G N N N N C A C
BsrGI  (4632)
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  (4307)
1 site
A G T A C T T C A T G A
PvuII  (4216)
1 site
C A G C T G G T C G A C
EcoNI  (3869)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
AvrII  (3865)
1 site
C C T A G G G G A T C C
AarI  (3819)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
AanI  (3792)
2 sites
T T A T A A A A T A T T
AsiSI  (5)
1 site
G C G A T C G C C G C T A G C G
PvuI  (5)
1 site
C G A T C G G C T A G C
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 ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BspDI  (977)
1 site
A T C G A T T A G C T A
ClaI  (977)
1 site
A T C G A T T A G C T A
BamHI  (986)
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.
StuI  (996)
1 site
A G G C C T T C C G G A
NheI  (1002)
1 site
G C T A G C C G A T C G
BmtI  (1006)
1 site
G C T A G C C G A T C G
AanI  (1152)
2 sites
T T A T A A A A T A T T
MfeI  (1181)
1 site
C A A T T G G T T A A C
PstI  (1518)
1 site
C T G C A G G A C G T C
SbfI  (1518)
1 site
C C T G C A G G G G A C G