pSF-pA-PromMCS-RLuc

Vector for low-background measurements of promoter activity using Renilla luciferase.

Sequence Author: Oxford Genetics

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PmeI (4649) RsrII (4342) BsrDI (4059) PflFI - Tth111I (3944) PmeI (3601) PpuMI - SanDI (3535) AscI (3444) FseI (3298) NaeI (3296) NgoMIV (3294) SwaI (3192) BspHI (3023) AlwNI (2719) BssSI (2476) AsiSI - PvuI (5) BglII (232) poly(A) signal ScaI (299) AarI (364) SalI (395) AccI (396) SpeI (403) TspMI - XmaI (411) SmaI (413) MCS PspOMI (419) ApaI (423) BstBI (428) BglII (435) EagI - NotI (477) HindIII (488) Eco53kI (498) SacI (500) EcoRI (504) Acc65I (516) KpnI (520) BstAPI (714) Bpu10I (1174) BseRI - BsgI (1463) stop codons BspDI - ClaI (1500) BamHI (1509) StuI (1519) NheI (1525) BmtI (1529) stop codons HpaI (1695) MfeI (1704) BglI (1864) PstI - SbfI (2041) 3' β-globin insulator BstEII (2147) PacI (2173) SwaI (2299) pSF-pA-PromMCS-RLuc 4768 bp
PmeI  (4649)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (4342)
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  (4059)
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  (3944)
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  (3944)
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  (3601)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (3535)
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  (3535)
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  (3444)
1 site
G G C G C G C C C C G C G C G G
FseI  (3298)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
NaeI  (3296)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (3294)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SwaI  (3192)
2 sites
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
BspHI  (3023)
1 site
T C A T G A A G T A C T
AlwNI  (2719)
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.
BssSI  (2476)
1 site
C A C G A G G T G C T C
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
ScaI  (299)
1 site
A G T A C T T C A T G A
AarI  (364)
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.
SalI  (395)
1 site
G T C G A C C A G C T G
AccI  (396)
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.
SpeI  (403)
1 site
A C T A G T T G A T C A
TspMI  (411)
1 site
C C C G G G G G G C C C
XmaI  (411)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (413)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PspOMI  (419)
1 site
G G G C C C C C C G G G
ApaI  (423)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BstBI  (428)
1 site
T T C G A A A A G C T T
BglII  (435)
2 sites
A G A T C T T C T A G A
EagI  (477)
1 site
C G G C C G G C C G G C
NotI  (477)
1 site
G C G G C C G C C G C C G G C G
HindIII  (488)
1 site
A A G C T T T T C G A A
Eco53kI  (498)
1 site
G A G C T C C T C G A G
SacI  (500)
1 site
G A G C T C C T C G A G
EcoRI  (504)
1 site
G A A T T C C T T A A G
Acc65I  (516)
1 site
G G T A C C C C A T G G
KpnI  (520)
1 site
G G T A C C C C A T G G
BstAPI  (714)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
Bpu10I  (1174)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BseRI  (1463)
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  (1463)
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  (1500)
1 site
A T C G A T T A G C T A
ClaI  (1500)
1 site
A T C G A T T A G C T A
BamHI  (1509)
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  (1519)
1 site
A G G C C T T C C G G A
NheI  (1525)
1 site
G C T A G C C G A T C G
BmtI  (1529)
1 site
G C T A G C C G A T C G
HpaI  (1695)
1 site
G T T A A C C A A T T G
MfeI  (1704)
1 site
C A A T T G G T T A A C
BglI  (1864)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
PstI  (2041)
1 site
C T G C A G G A C G T C
SbfI  (2041)
1 site
C C T G C A