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Plasmid Files

phRL-CMV

Mammalian vector for strong constitutive expression of humanized Renilla luciferase.

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phRL-CMV Sequence and MapphRL-CMV.dna
Map and Sequence File   
Sequence Author:  Promega
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 HaeII (3828) PspFI (3768) BseYI (3764) AlwNI (3659) AhdI (3180) BsrFI (3095) FspI (2957) PvuI (2811) BamHI (2257) BspDI - ClaI (2250) MfeI (2164) HpaI (2155) PsiI (2135) ApoI (2100) EagI - NotI (2012) BglII (1) BsrGI (102) SpeI (158) NdeI (393) BtgZI (493) BsaAI - SnaBI (499) Eco53kI (725) SacI (727) HindIII (754) PstI (836) BfuAI - BspMI (850) BbsI (934) NheI (1058) BmtI (1062) AvaI - BsoBI (1089) BmeT110I (1090) DraIII (1256) NsiI (1287) PfoI * (1348) EcoRV (1553) Bsu36I (1654) NruI (1729) PflFI - Tth111I (1764) PasI (1842) EcoO109I (1913) XbaI (2005) phRL-CMV 4079 bp
HaeII  (3828)
1 site
R G C G C Y Y C G C G R
PspFI  (3768)
1 site
C C C A G C G G G T C G
BseYI  (3764)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
AlwNI  (3659)
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  (3180)
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  (3095)
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.
FspI  (2957)
1 site
T G C G C A A C G C G T
PvuI  (2811)
1 site
C G A T C G G C T A G C
BamHI  (2257)
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  (2250)
1 site
A T C G A T T A G C T A
ClaI  (2250)
1 site
A T C G A T T A G C T A
MfeI  (2164)
1 site
C A A T T G G T T A A C
HpaI  (2155)
1 site
G T T A A C C A A T T G
PsiI  (2135)
1 site
T T A T A A A A T A T T
ApoI  (2100)
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.
EagI  (2012)
1 site
C G G C C G G C C G G C
NotI  (2012)
1 site
G C G G C C G C C G C C G G C G
BglII  (1)
1 site
A G A T C T T C T A G A
BsrGI  (102)
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.
SpeI  (158)
1 site
A C T A G T T G A T C A
NdeI  (393)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
BtgZI  (493)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsaAI  (499)
1 site
Y A C G T R R T G C A Y
SnaBI  (499)
1 site
T A C G T A A T G C A T
Eco53kI  (725)
1 site
G A G C T C C T C G A G
SacI  (727)
1 site
G A G C T C C T C G A G
HindIII  (754)
1 site
A A G C T T T T C G A A
PstI  (836)
1 site
C T G C A G G A C G T C
BfuAI  (850)
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  (850)
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  (934)
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  (1058)
1 site
G C T A G C C G A T C G
BmtI  (1062)
1 site
G C T A G C C G A T C G
AvaI  (1089)
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  (1089)
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  (1090)
1 site
C Y C G R G G R G C Y C
DraIII  (1256)
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.
NsiI  (1287)
1 site
A T G C A T T A C G T A
PfoI  (1348)
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  (1553)
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  (1654)
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  (1729)
1 site
T C G C G A A G C G C T
PflFI  (1764)
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  (1764)
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.
PasI  (1842)
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  (1913)
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  (2005)
1 site
T C T A G A A G A T C T
hRluc
1068 .. 2003  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
1068 .. 2003  =  936 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
AmpR
2393 .. 3253  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2393 .. 2461  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2393 .. 3253  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2462 .. 3253  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2393 .. 3253  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3424 .. 4012  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3424 .. 4012  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
220 .. 523  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
220 .. 523  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
524 .. 727  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
524 .. 727  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
863 .. 995  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
863 .. 995  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
2034 .. 2155  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2034 .. 2155  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2288 .. 2392  =  105 bp
AmpR promoter
2288 .. 2392  =  105 bp
T7 promoter
1040 .. 1058  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1040 .. 1058  =  19 bp
promoter for bacteriophage T7 RNA polymerase
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