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Plasmid Files

pSF-CMV-FMDV-FLuc

Mammalian vector for co-expressing a gene together with firefly luciferase expressed from the foot-and-mouth disease virus IRES.

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pSF-CMV-FMDV-FLuc Sequence and MappSF-CMV-FMDV-FLuc.dna
Map and Sequence File   
Sequence Author:  Oxford Genetics
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 PmeI (6250) RsrII (5943) BsrDI (5660) PflFI - Tth111I (5545) PmeI (5202) SanDI (5136) AscI - BssHII (5045) FseI (4899) SwaI (4793) AlwNI (4320) ApaLI (4218) SwaI (3900) PacI (3774) 3' β- globin insulator PstI - SbfI (3642) MfeI (3305) AanI - AanI - PsiI (3276) stop codons AsiSI - PvuI (5) BglII (232) SnaBI (566) BglII (816) NotI (858) Eco53kI (879) SacI (881) EcoRI (885) NcoI (905) EcoRV (921) AbsI - PaeR7I - PspXI - XhoI (928) BsgI (940) BspDI - ClaI (977) EcoNI (1011) PspOMI (1226) ApaI (1230) PflMI (1308) AhdI (1438) BbsI (1531) PvuII (1547) ScaI (1638) BsrGI (1963) FspI (2295) KasI (2420) NarI (2421) SfoI (2422) PluTI (2424) BlpI (2427) AgeI (2604) DraIII (2619) SexAI * (2812) BpmI (2853) NheI (3126) BmtI (3130) pSF-CMV-FMDV-FLuc 6369 bp
PmeI  (6250)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (5943)
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  (5660)
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  (5545)
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  (5545)
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  (5202)
2 sites
G T T T A A A C C A A A T T T G
SanDI  (5136)
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  (5045)
1 site
G G C G C G C C C C G C G C G G
BssHII  (5045)
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  (4899)
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  (4793)
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  (4320)
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  (4218)
1 site
G T G C A C C A C G T G
SwaI  (3900)
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  (3774)
1 site
T T A A T T A A A A T T A A T T
PstI  (3642)
1 site
C T G C A G G A C G T C
SbfI  (3642)
1 site
C C T G C A G G G G A C G T C C
MfeI  (3305)
1 site
C A A T T G G T T A A C
AanI  (3276)
1 site
T T A T A A A A T A T T
AanI  (3276)
1 site
T T A T A A A A T A T T
PsiI  (3276)
1 site
T T A T A A A A T A T T
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
NotI  (858)
1 site
G C G G C C G C C G C C G G C G
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
NcoI  (905)
1 site
C C A T G G G G T A C C
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
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
EcoNI  (1011)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PspOMI  (1226)
1 site
G G G C C C C C C G G G
ApaI  (1230)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PflMI  (1308)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
AhdI  (1438)
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.
BbsI  (1531)
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  (1547)
1 site
C A G C T G G T C G A C
ScaI  (1638)
1 site
A G T A C T T C A T G A
BsrGI  (1963)
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.
FspI  (2295)
1 site
T G C G C A A C G C G T
KasI  (2420)
1 site
G G C G C C C C G C G G
NarI  (2421)
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  (2422)
1 site
G G C G C C C C G C G G
PluTI  (2424)
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  (2427)
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  (2604)
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  (2619)
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  (2812)
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  (2853)
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.
NheI  (3126)
1 site
G C T A G C C G A T C G
BmtI  (3130)
1 site
G C T A G C C G A T C G
luciferase
1473 .. 3125  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
1473 .. 3125  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
NeoR/KanR
5299 .. 6093  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
5299 .. 6093  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
3965 .. 4553  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3965 .. 4553  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
1012 .. 1457  =  446 bp
internal ribosome entry site (IRES) of the
foot-and-mouth disease virus (FMDV)
IRES
1012 .. 1457  =  446 bp
internal ribosome entry site (IRES) of the
foot-and-mouth disease virus (FMDV)
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
3487 .. 3623  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
3487 .. 3623  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
6106 .. 6242  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
6106 .. 6242  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
3175 .. 3296  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3175 .. 3296  =  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
3668 .. 3739  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
3' β-globin insulator
3668 .. 3739  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
T7 terminator
3411 .. 3457  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
3411 .. 3457  =  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
3133 .. 3143  =  11 bp
stop codons in all three reading frames
stop codons
3133 .. 3143  =  11 bp
stop codons in all three reading frames
RBS
5286 .. 5291  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
5286 .. 5291  =  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
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