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Plasmid Files

pRL-SV40

Mammalian co-reporter vector for the strong constitutive expression of wild-type Renilla luciferase.

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pRL-SV40 Sequence and MappRL-SV40.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (1) HaeII (3454) AlwNI (3285) AhdI (2806) BpmI (2737) BsrFI (2721) PvuI (2437) BsaHI (2266) SspI (2001) BamHI (1883) BsaBI * (1882) BspDI - ClaI (1876) Acc65I (54) KpnI (58) PvuII (80) SexAI * (171) SfiI (357) BseRI (400) StuI (403) AvrII (404) HindIII (420) PstI (462) BfuAI - BspMI (476) NheI (684) BmtI (688) BstBI (700) AflIII - PciI (876) XcmI (1343) BsrGI (1392) BsaAI (1426) XbaI (1631) EagI - NotI (1638) HincII - HpaI (1781) MfeI (1790) pRL-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.
BpmI  (2737)
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.
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.
PvuI  (2437)
1 site
C G A T C G G C T A G C
BsaHI  (2266)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°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.
BsaBI  (1882)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
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.
BseRI  (400)
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.
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.
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
BstBI  (700)
1 site
T T C G A A A A G C T T
AflIII  (876)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (876)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
XcmI  (1343)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BsrGI  (1392)
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.
BsaAI  (1426)
1 site
Y A C G T R R T G C A Y
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
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
Rluc
694 .. 1629  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)
Rluc
694 .. 1629  =  936 bp
311 amino acids  =  36.0 kDa
Product: luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)
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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