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Plasmid Files

phRL-SV40

Mammalian vector for strong constitutive expression of humanized Renilla luciferase.

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phRL-SV40 Sequence and MapphRL-SV40.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (1) HaeII (3454) AlwNI (3285) AhdI (2806) BsrFI (2721) AseI (2631) PvuI (2437) SspI (2001) BamHI (1883) BspDI - ClaI (1876) Acc65I (54) KpnI (58) PvuII (80) SexAI * (171) SfiI (357) StuI (403) AvrII (404) HindIII (420) PstI (462) BfuAI - BspMI (476) BbsI (560) NheI (684) BmtI (688) AvaI - BsoBI (715) BmeT110I (716) DraIII (882) PfoI * (974) EcoRV (1179) Bsu36I (1280) NruI (1355) ZraI (1389) PflFI - Tth111I (1390) AatII (1391) PasI (1468) EcoO109I (1539) XbaI (1631) EagI - NotI (1638) ApoI (1726) PsiI (1761) HincII - HpaI (1781) MfeI (1790) phRL-SV40 3705 bp
BglII  (1)
1 site
A G A T C T T C T A G A
HaeII  (3454)
1 site
R G C G C Y Y C G C G R
AlwNI  (3285)
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.
AhdI  (2806)
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.
BsrFI  (2721)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
AseI  (2631)
1 site
A T T A A T T A A T T A
PvuI  (2437)
1 site
C G A T C G G C T A G C
SspI  (2001)
1 site
A A T A T T T T A T A A
BamHI  (1883)
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.
BspDI  (1876)
1 site
A T C G A T T A G C T A
ClaI  (1876)
1 site
A T C G A T T A G C T A
Acc65I  (54)
1 site
G G T A C C C C A T G G
KpnI  (58)
1 site
G G T A C C C C A T G G
PvuII  (80)
1 site
C A G C T G G T C G A C
SexAI  (171)
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.
SfiI  (357)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
StuI  (403)
1 site
A G G C C T T C C G G A
AvrII  (404)
1 site
C C T A G G G G A T C C
HindIII  (420)
1 site
A A G C T T T T C G A A
PstI  (462)
1 site
C T G C A G G A C G T C
BfuAI  (476)
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  (476)
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.
BbsI  (560)
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.
NheI  (684)
1 site
G C T A G C C G A T C G
BmtI  (688)
1 site
G C T A G C C G A T C G
AvaI  (715)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (715)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
BmeT110I  (716)
1 site
C Y C G R G G R G C Y C
DraIII  (882)
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.
PfoI  (974)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
EcoRV  (1179)
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.
Bsu36I  (1280)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
NruI  (1355)
1 site
T C G C G A A G C G C T
ZraI  (1389)
1 site
G A C G T C C T G C A G
PflFI  (1390)
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  (1390)
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.
AatII  (1391)
1 site
G A C G T C C T G C A G
PasI  (1468)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoO109I  (1539)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
XbaI  (1631)
1 site
T C T A G A A G A T C T
EagI  (1638)
1 site
C G G C C G G C C G G C
NotI  (1638)
1 site
G C G G C C G C C G C C G G C G
ApoI  (1726)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
PsiI  (1761)
1 site
T T A T A A A A T A T T
HincII  (1781)
1 site
G T Y R A C C A R Y T G
HpaI  (1781)
1 site
G T T A A C C A A T T G
MfeI  (1790)
1 site
C A A T T G G T T A A C
hRluc
694 .. 1629  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
694 .. 1629  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
AmpR
2019 .. 2879  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2019 .. 2087  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2019 .. 2879  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2088 .. 2879  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2019 .. 2879  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3050 .. 3638  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3050 .. 3638  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 promoter
62 .. 419  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
62 .. 419  =  358 bp
SV40 enhancer and early promoter
chimeric intron
489 .. 621  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
489 .. 621  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
1660 .. 1781  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1660 .. 1781  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1914 .. 2018  =  105 bp
AmpR promoter
1914 .. 2018  =  105 bp
T7 promoter
666 .. 684  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
666 .. 684  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
270 .. 405  =  136 bp
SV40 origin of replication
SV40 ori
270 .. 405  =  136 bp
SV40 origin of replication
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