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
NotI  (498)
1 site
G C G G C C G C C G C C G G C G
HindIII  (509)
1 site
A A G C T T T T C G A A
Eco53kI  (519)
1 site
G A G C T C C T C G A G
SacI  (521)
1 site
G A G C T C C T C G A G
EcoRI  (525)
1 site
G A A T T C C T T A A G
Acc65I  (537)
1 site
G G T A C C C C A T G G
KpnI  (541)
1 site
G G T A C C C C A T G G
NcoI  (545)
1 site
C C A T G G G G T A C C
BbsI  (605)
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.
PvuII  (621)
1 site
C A G C T G G T C G A C
BsrGI  (1037)
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.
AleI  (1045)
1 site
C A C N N N N G T G G T G N N N N C A C
FspI  (1369)
1 site
T G C G C A A C G C G T
KasI  (1494)
1 site
G G C G C C C C G C G G
NarI  (1495)
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.
SfoI  (1496)
1 site
G G C G C C C C G C G G
PluTI  (1498)
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.
BlpI  (1501)
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.
AgeI  (1678)
1 site
A C C G G T T G G C C A
DraIII  (1693)
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.
SexAI  (1886)
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.
BpmI  (1927)
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.
XbaI  (2195)
1 site
T C T A G A A G A T C T
BseRI  (2198)
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  (2198)
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  (2235)
1 site
A T C G A T T A G C T A
ClaI  (2235)
1 site
A T C G A T T A G C T A
BamHI  (2244)
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  (2254)
1 site
A G G C C T T C C G G A
NheI  (2260)
1 site
G C T A G C C G A T C G
BmtI  (2264)
1 site
G C T A G C C G A T C G
BtsI  (2356)
1 site
G C A G T G N N C G T C A C
PsiI  (2410)
1 site
T T A T A A A A T A T T
MfeI  (2439)
1 site
C A A T T G G T T A A C
BglI  (2599)
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.
SbfI  (2776)
1 site
C C T G C A G G G G A C G T C C
luciferase
547 .. 2199  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
547 .. 2199  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
NeoR/KanR
4433 .. 5227  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4433 .. 5227  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3099 .. 3687  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3099 .. 3687  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rrnG terminator
2621 .. 2757  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
2621 .. 2757  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5240 .. 5376  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5240 .. 5376  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
2309 .. 2430  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2309 .. 2430  =  122 bp
SV40 polyadenylation signal
pause site
245 .. 336  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
245 .. 336  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus (Farrell et al., 2002)
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2802 .. 2873  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2802 .. 2873  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
Mini-TK promoter
379 .. 441  =  63 bp
minimal herpes simplex virus (HSV) thymidine kinase promoter
Mini-TK promoter
379 .. 441  =  63 bp
minimal herpes simplex virus (HSV) thymidine kinase promoter
T7 terminator
2545 .. 2591  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2545 .. 2591  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
MCS
337 .. 374  =  38 bp
multiple cloning site
MCS
337 .. 374  =  38 bp
multiple cloning site
stop codons
2219 .. 2229  =  11 bp
stop codons in all three reading frames
stop codons
2219 .. 2229  =  11 bp
stop codons in all three reading frames
stop codons
2267 .. 2277  =  11 bp
stop codons in all three reading frames
stop codons
2267 .. 2277  =  11 bp
stop codons in all three reading frames
RBS
533 .. 538  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
533 .. 538  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4420 .. 4425  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4420 .. 4425  =  6 bp
Shine-Dalgarno ribosome binding site
Kozak sequence
543 .. 549  =  7 bp
Kozak sequence
543 .. 549  =  7 bp
ORF:  547 .. 2199  =  1653 bp
ORF:  550 amino acids  =  60.6 kDa
ORF:  4433 .. 5227  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2646 .. 3077  =  432 bp
ORF:  143 amino acids  =  16.3 kDa
ORF:  4605 .. 4991  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  5265 .. 196  =  435 bp
ORF:  144 amino acids  =  16.1 kDa
ORF:  529 .. 774  =  246 bp
ORF:  81 amino acids  =  8.8 kDa
ORF:  1543 .. 1803  =  261 bp
ORF:  86 amino acids  =  8.9 kDa
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