pSF-CMV-PGK-RLuc

Dual-promoter mammalian vector for strong constitutive expression of a gene together with Renilla luciferase.

Sequence Author: Oxford Genetics

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PmeI (5561) RsrII (5254) PflFI - Tth111I (4856) PmeI (4513) SanDI (4447) AscI (4356) FseI (4210) NaeI (4208) NgoMIV (4206) SwaI (4104) BspHI (3935) SwaI (3211) PacI (3085) BstEII (3059) 3' β-globin insulator PstI - SbfI (2953) AsiSI - PvuI (5) BglII (232) CMV enhancer 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) SphI (1027) BpmI (1039) AgeI (1098) BsmBI - SpeI (1233) BlpI (1261) BamHI (1488) BstAPI (1686) AanI (1790) NheI (2437) BmtI (2441) stop codons AanI (2587) HpaI (2607) MfeI (2616) pSF-CMV-PGK-RLuc 5680 bp
PmeI  (5561)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (5254)
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.
PflFI  (4856)
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  (4856)
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  (4513)
2 sites
G T T T A A A C C A A A T T T G
SanDI  (4447)
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  (4356)
1 site
G G C G C G C C C C G C G C G G
FseI  (4210)
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  (4208)
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  (4206)
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  (4104)
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  (3935)
1 site
T C A T G A A G T A C T
SwaI  (3211)
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  (3085)
1 site
T T A A T T A A A A T T A A T T
BstEII  (3059)
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.
PstI  (2953)
1 site
C T G C A G G A C G T C
SbfI  (2953)
1 site
C C T G C A G G G G A C G T C 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
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
SphI  (1027)
1 site
G C A T G C C G T A C G
BpmI  (1039)
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.
AgeI  (1098)
1 site
A C C G G T T G G C C A
BsmBI  (1233)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
SpeI  (1233)
1 site
A C T A G T T G A T C A
BlpI  (1261)
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.
BamHI  (1488)
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.
BstAPI  (1686)
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.
AanI  (1790)
2 sites
T T A T A A A A T A T T
NheI  (2437)
1 site
G C T A G C C G A T C G
BmtI  (2441)
1 site
G C T A G C C G A T C G
AanI  (2587)
2 sites
T T A T A A A A T A T T
HpaI  (2607)
1 site
G T T A A C C A A T T G
MfeI  (2616)
1 site
C A A T T G G T T A A C
RLuc
1498 .. 2436  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
RLuc
1498 .. 2436  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
NeoR/KanR
4610 .. 5404  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4610 .. 5404  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3276 .. 3864  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3276 .. 3864  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
988 .. 1487  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
988 .. 1487  =  500 bp
mouse phosphoglycerate kinase 1 promoter
CMV enhancer
287 .. 590  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
287 .. 590  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
591 .. 794  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
591 .. 794  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
rrnG terminator
2798 .. 2934  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
2798 .. 2934  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5417 .. 5553  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5417 .. 5553  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
2486 .. 2607  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2486 .. 2607  =  122 bp
SV40 polyadenylation signal
MCS
857 .. 942  =  86 bp
multiple cloning site
MCS
857 .. 942  =  86 bp
multiple cloning site
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
2979 .. 3050  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2979 .. 3050  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
T7 terminator
2722 .. 2768  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2722 .. 2768  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
stop codons
961 .. 971  =  11 bp
stop codons in all three reading frames
stop codons
961 .. 971  =  11 bp
stop codons in all three reading frames
stop codons
2444 .. 2454  =  11 bp
stop codons in all three reading frames
stop codons
2444 .. 2454  =  11 bp
stop codons in all three reading frames
RBS
4597 .. 4602  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4597 .. 4602  =  6 bp
Shine-Dalgarno ribosome binding site
Kozak sequence
903 .. 909  =  7 bp
Kozak sequence
903 .. 909  =  7 bp
RBS
893 .. 898  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
893 .. 898  =  6 bp
Shine-Dalgarno ribosome binding site
ATG
907 .. 909  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
907 .. 909  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  1498 .. 2436  =  939 bp
ORF:  312 amino acids  =  36.1 kDa
ORF:  1268 .. 1501  =  234 bp
ORF:  77 amino acids  =  8.2 kDa
ORF:  4610 .. 5404  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2823 .. 3254  =  432 bp
ORF:  143 amino acids  =  16.3 kDa
ORF:  4782 .. 5168  =  387 bp
ORF:  128 amino acids  =  14.7 kDa
ORF:  5442 .. 196  =  435 bp
ORF:  144 amino acids  =  16.1 kDa
ORF:  1378 .. 2352  =  975 bp
ORF:  324 amino acids  =  38.5 kDa
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