pSF-MinProm-FLuc

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

Sequence Author: Oxford Genetics

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AsiSI - PvuI (5) PmeI (5384) RsrII (5077) BsrDI (4794) PflFI - Tth111I (4679) PmeI (4336) PpuMI - SanDI (4270) AscI - BssHII (4179) FseI (4033) SwaI (3927) AlwNI (3454) ApaLI (3352) SwaI (3034) PacI (2908) BstEII (2882) 3' β-globin insulator 5' β-globin insulator BglII (232) AarI (306) SalI (337) AccI (338) SpeI (345) TspMI - XmaI (353) SmaI (355) MCS PspOMI (361) ApaI (365) BstBI (370) MluI (395) BglII (456) EagI - NotI (498) HindIII (509) Eco53kI (519) SacI (521) EcoRI (525) Acc65I (537) KpnI (541) NcoI (545) BbsI (605) PvuII (621) BsrGI (1037) AleI (1045) FspI (1369) KasI (1494) NarI (1495) SfoI (1496) PluTI (1498) BlpI (1501) AgeI (1678) DraIII (1693) SexAI * (1886) BpmI (1927) XbaI (2195) BseRI - BsgI (2198) stop codons BspDI - ClaI (2235) BamHI (2244) StuI (2254) NheI (2260) BmtI (2264) stop codons BtsI (2356) PsiI (2410) MfeI (2439) BglI (2599) SbfI (2776) pSF-MinProm-FLuc 5503 bp
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
PmeI  (5384)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (5077)
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  (4794)
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  (4679)
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  (4679)
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  (4336)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (4270)
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  (4270)
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  (4179)
1 site
G G C G C G C C C C G C G C G G
BssHII  (4179)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
FseI  (4033)
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.
SwaI  (3927)
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.
AlwNI  (3454)
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.
ApaLI  (3352)
1 site
G T G C A C C A C G T G
SwaI  (3034)
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.
PacI  (2908)
1 site
T T A A T T A A A A T T A A T T
BstEII  (2882)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BglII  (232)
2 sites
A G A T C T T C T A G A
AarI  (306)
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  (337)
1 site
G T C G A C C A G C T G
AccI  (338)
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  (345)
1 site
A C T A G T T G A T C A
TspMI  (353)
1 site
C C C G G G G G G C C C
XmaI  (353)
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  (355)
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  (361)
1 site
G G G C C C C C C G G G
ApaI  (365)
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  (370)
1 site
T T C G A A A A G C T T
MluI  (395)
1 site
A C G C G T T G C G C A
BglII  (456)
2 sites
A G A T C T T C T A G A
EagI  (498)
1 site
C G G C C G G C C G G C