pSF-CMV-RLuc

Mammalian vector for strong constitutive expression of Renilla luciferase.

Sequence Author: Oxford Genetics

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BtgI (5157) PmeI (5103) AhdI (4866) BsaI (4800) BpmI (4797) PmeI (3982) PpuMI - SanDI (3916) AscI (3825) FseI (3679) NaeI (3677) NgoMIV (3675) SwaI (3573) BspHI (3404) AlwNI (3100) BssSI (2857) DrdI (2792) SwaI (2680) AsiSI - PvuI (5) BglII (232) CMV enhancer SnaBI (566) BglII (816) EagI - NotI (858) HindIII (869) Eco53kI (879) BanII - SacI (881) EcoRI (885) RBS Acc65I (897) KpnI (901) BstAPI (1095) BfuAI - BspMI (1241) Bpu10I (1555) BseRI - BsgI (1844) stop codons BspDI - ClaI (1881) BamHI (1890) StuI (1900) NheI (1906) BmtI (1910) stop codons HpaI (2076) MfeI (2085) PstI - SbfI (2422) 3' β-globin insulator PacI (2554) pSF-CMV-RLuc 5222 bp
BtgI  (5157)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
PmeI  (5103)
2 sites
G T T T A A A C C A A A T T T G
AhdI  (4866)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsaI  (4800)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BpmI  (4797)
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.
PmeI  (3982)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (3916)
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  (3916)
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  (3825)
1 site
G G C G C G C C C C G C G C G G
FseI  (3679)
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  (3677)
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  (3675)
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  (3573)
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  (3404)
1 site
T C A T G A A G T A C T
AlwNI  (3100)
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  (2857)
1 site
C A C G A G G T G C T C
DrdI  (2792)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
SwaI  (2680)
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
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
BanII  (881)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
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
Acc65I  (897)
1 site
G G T A C C C C A T G G
KpnI  (901)
1 site
G G T A C C C C A T G G
BstAPI  (1095)
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.
BfuAI  (1241)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1241)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
Bpu10I  (1555)
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  (1844)
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  (1844)
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  (1881)
1 site
A T C G A T T A G C T A
ClaI  (1881)
1 site
A T C G A T T A G C T A
BamHI  (1890)
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  (1900)
1 site
A G G C C T T C C G G A
NheI  (1906)
1 site
G C T A G C C G A T C G
BmtI  (1910)
1 site
G C T A G C C G A T C G
HpaI  (2076)
1 site
G T T A A C C A A T T G
MfeI  (2085)
1 site
C A A T T G G T T A A C
PstI  (2422)
1 site
C T G C A G G A C G T C
SbfI  (2422)
1 site
C C T G C A G G G G A C G T C C
PacI  (2554)
1 site
T T A A T T A A A A T T A A T T
RLuc
907 .. 1845  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
RLuc
907 .. 1845  =  939 bp
312 amino acids  =  36.1 kDa
Product: Renilla luciferase
codon-optimized synthetic gene
AmpR
4079 .. 4939  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4079 .. 4147  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4079 .. 4939  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4148 .. 4939  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4079 .. 4939  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2745 .. 3333  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2745 .. 3333  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
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
2267 .. 2403  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
2267 .. 2403  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
4952 .. 5088  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
4952 .. 5088  =  137 bp
transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
1955 .. 2076  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1955 .. 2076  =  122 bp
SV40 polyadenylation signal
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
2448 .. 2519  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
3' β-globin insulator
2448 .. 2519  =  72 bp
insulator downstream of the human β-globin locus (Farrell et al., 2002)
T7 terminator
2191 .. 2237  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2191 .. 2237  =  47 bp
transcription terminator for bacteriophage T7 RNA polymerase
stop codons
1865 .. 1875  =  11 bp
stop codons in all three reading frames
stop codons
1865 .. 1875  =  11 bp
stop codons in all three reading frames
stop codons
1913 .. 1923  =  11 bp
stop codons in all three reading frames
stop codons
1913 .. 1923  =  11 bp
stop codons in all three reading frames
RBS
893 .. 898  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
893 .. 898  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4066 .. 4071  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4066 .. 4071  =  6 bp
Shine-Dalgarno ribosome binding site
Kozak sequence
903 .. 909  =  7 bp
Kozak sequence
903 .. 909  =  7 bp
ORF:  907 .. 1845  =  939 bp
ORF:  312 amino acids  =  36.1 kDa
ORF:  4079 .. 4939  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4970 .. 59  =  312 bp
ORF:  103 amino acids  =  11.6 kDa
ORF:  2292 .. 2723  =  432 bp
ORF:  143 amino acids  =  16.3 kDa
ORF:  889 .. 1761  =  873 bp
ORF:  290 amino acids  =  34.7 kDa
ORF:  4543 .. 4809  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
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