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Plasmid Files

pSF-CMV-FLuc

Mammalian vector for strong constitutive expression of firefly luciferase.

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pSF-CMV-FLuc Sequence and MappSF-CMV-FLuc.dna
Map and Sequence File   
Sequence Author:  Oxford Genetics
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 AsiSI - PvuI (5) PmeI (5817) AhdI (5580) BsaI (5514) PmeI (4696) PpuMI - SanDI (4630) AscI - BssHII (4539) FseI (4393) SwaI (4287) AlwNI (3814) ApaLI (3712) SwaI (3394) PacI (3268) 3' β- globin insulator PstI - SbfI (3136) 5' β- globin insulator BglII (232) CMV enhancer SnaBI (566) BglII (816) EagI - NotI (858) HindIII (869) Eco53kI (879) BanII - SacI (881) EcoRI (885) Acc65I (897) KpnI (901) NcoI (905) BbsI (965) PvuII (981) ScaI (1072) BsrGI (1397) AleI (1405) FspI (1729) KasI (1854) NarI (1855) SfoI (1856) PluTI (1858) BlpI (1861) AgeI (2038) DraIII (2053) SexAI * (2246) XbaI (2555) BseRI - BsgI (2558) stop codons BspDI - ClaI (2595) BamHI (2604) StuI (2614) NheI (2620) BmtI (2624) stop codons PsiI (2770) MfeI (2799) pSF-CMV-FLuc 5936 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  (5817)
2 sites
G T T T A A A C C A A A T T T G
AhdI  (5580)
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  (5514)
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.
PmeI  (4696)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (4630)
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  (4630)
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  (4539)
1 site
G G C G C G C C C C G C G C G G
BssHII  (4539)
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  (4393)
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  (4287)
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  (3814)
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  (3712)
1 site
G T G C A C C A C G T G
SwaI  (3394)
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  (3268)
1 site
T T A A T T A A A A T T A A T T
PstI  (3136)
1 site
C T G C A G G A C G T C
SbfI  (3136)
1 site
C C T G C A G G G G A C G T C 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
NcoI  (905)
1 site
C C A T G G G G T A C C
BbsI  (965)
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  (981)
1 site
C A G C T G G T C G A C
ScaI  (1072)
1 site
A G T A C T T C A T G A
BsrGI  (1397)
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  (1405)
1 site
C A C N N N N G T G G T G N N N N C A C
FspI  (1729)
1 site
T G C G C A A C G C G T
KasI  (1854)
1 site
G G C G C C C C G C G G
NarI  (1855)
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  (1856)
1 site
G G C G C C C C G C G G
PluTI  (1858)
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  (1861)
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  (2038)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
DraIII  (2053)
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  (2246)
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.
XbaI  (2555)
1 site
T C T A G A A G A T C T
BseRI  (2558)
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  (2558)
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  (2595)
1 site
A T C G A T T A G C T A
ClaI  (2595)
1 site
A T C G A T T A G C T A
BamHI  (2604)
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  (2614)
1 site
A G G C C T T C C G G A
NheI  (2620)
1 site
G C T A G C C G A T C G
BmtI  (2624)
1 site
G C T A G C C G A T C G
PsiI  (2770)
1 site
T T A T A A A A T A T T
MfeI  (2799)
1 site
C A A T T G G T T A A C
luciferase
907 .. 2559  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
907 .. 2559  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
AmpR
4793 .. 5653  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4793 .. 4861  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4793 .. 5653  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4862 .. 5653  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4793 .. 5653  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3459 .. 4047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3459 .. 4047  =  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
2981 .. 3117  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
2981 .. 3117  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5666 .. 5802  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5666 .. 5802  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
2669 .. 2790  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2669 .. 2790  =  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
3162 .. 3233  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
3' β-globin insulator
3162 .. 3233  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
T7 terminator
2905 .. 2951  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
2905 .. 2951  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
stop codons
2579 .. 2589  =  11 bp
stop codons in all three reading frames
stop codons
2579 .. 2589  =  11 bp
stop codons in all three reading frames
stop codons
2627 .. 2637  =  11 bp
stop codons in all three reading frames
stop codons
2627 .. 2637  =  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
4780 .. 4785  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4780 .. 4785  =  6 bp
Shine-Dalgarno ribosome binding site
Kozak sequence
903 .. 909  =  7 bp
Kozak sequence
903 .. 909  =  7 bp
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