pSF-pA-MinProm-RLuc

Vector encoding a minimal promoter and Renilla luciferase, for measuring the activity of transcriptional regulatory elements.

Sequence Author: Oxford Genetics

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PmeI (4826) RsrII (4519) BsrDI (4236) PflFI - Tth111I (4121) PmeI (3778) PpuMI - SanDI (3712) AscI (3621) FseI (3475) NaeI (3473) NgoMIV (3471) SwaI (3369) BspHI (3200) AlwNI (2896) BssSI (2653) SwaI (2476) AsiSI - PvuI (5) BglII (232) SexAI * (249) NruI (318) PshAI (324) poly(A) signal AarI (462) SalI (493) AccI (494) SpeI (501) TspMI - XmaI (509) SmaI (511) PspOMI (517) ApaI (521) BstBI (526) BglII (612) EagI - NotI (654) HindIII (665) Eco53kI (675) SacI (677) EcoRI (681) Acc65I (693) KpnI (697) BstAPI (891) Bpu10I (1351) BseRI - BsgI (1640) stop codons BspDI - ClaI (1677) BamHI (1686) StuI (1696) NheI (1702) BmtI (1706) stop codons HpaI (1872) MfeI (1881) BglI (2041) SbfI (2218) 3' β-globin insulator BstEII (2324) PacI (2350) pSF-pA-MinProm-RLuc 4945 bp
PmeI  (4826)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (4519)
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  (4236)
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  (4121)
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  (4121)
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  (3778)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (3712)
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  (3712)
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  (3621)
1 site
G G C G C G C C C C G C G C G G
FseI  (3475)
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  (3473)
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  (3471)
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  (3369)
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  (3200)
1 site
T C A T G A A G T A C T
AlwNI  (2896)
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  (2653)
1 site
C A C G A G G T G C T C
SwaI  (2476)
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.
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
SexAI  (249)
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.
NruI  (318)
1 site
T C G C G A A G C G C T
PshAI  (324)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
AarI  (462)
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  (493)
1 site
G T C G A C C A G C T G
AccI  (494)
1 site