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Plasmid Files

pGL4.51[luc2/CMV/Neo]

Vector for creating stable cell lines expressing firefly luciferase under control of the CMV promoter.

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pGL4.51[luc2 CMV Neo].dna
Map and Sequence File:    Download    Open   
Sequence Author:  Promega
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RVprimer3 (6307 .. 6326) BsmBI (6155) BstZ17I (5823) SacII (5707) PvuI (5683) Bsu36I (5669) AhdI (5313) BstEII (5238) BstXI - PstI (5235) NotI (5211) ApaLI (4705) BciVI (4594) AflIII - PciI (4391) RVprimer4 (4192 .. 4211) SalI (4141) BstBI (4127) EcoNI (3732) StuI (3211) AarI (6328) SfiI (8) AseI (173) NdeI (400) BsaAI - SnaBI (506) Eco53kI (732) SacI (734) SfiI (818) MreI (922) DraIII (2005) BpmI (2239) ApoI (2621) PsiI (2656) MfeI (2685) BamHI (2778) pGL4.51[luc2/CMV/Neo] 6358 bp
BsmBI  (6155)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BstZ17I  (5823)
1 site
G T A T A C C A T A T G
SacII  (5707)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PvuI  (5683)
1 site
C G A T C G G C T A G C
Bsu36I  (5669)
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.
AhdI  (5313)
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.
BstEII  (5238)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BstXI  (5235)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
PstI  (5235)
1 site
C T G C A G G A C G T C
NotI  (5211)
1 site
G C G G C C G C C G C C G G C G
ApaLI  (4705)
1 site
G T G C A C C A C G T G
BciVI  (4594)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
AflIII  (4391)
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  (4391)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (4141)
1 site
G T C G A C C A G C T G
BstBI  (4127)
1 site
T T C G A A A A G C T T
EcoNI  (3732)
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.
StuI  (3211)
1 site
A G G C C T T C C G G A
AarI  (6328)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
SfiI  (8)
2 sites
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.
AseI  (173)
1 site
A T T A A T T A A T T A
NdeI  (400)
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.
BsaAI  (506)
1 site
Y A C G T R R T G C A Y
SnaBI  (506)
1 site
T A C G T A A T G C A T
Eco53kI  (732)
1 site
G A G C T C C T C G A G
SacI  (734)
1 site
G A G C T C C T C G A G
SfiI  (818)
2 sites
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.
MreI  (922)
1 site
C G C C G G C G G C G G C C G C
DraIII  (2005)
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.
BpmI  (2239)
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.
ApoI  (2621)
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  (2656)
1 site
T T A T A A A A T A T T
MfeI  (2685)
1 site
C A A T T G G T T A A C
BamHI  (2778)
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.
RVprimer3
20-mer  /  50% GC
1 binding site
6307 .. 6326  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
4192 .. 4211  =  20 annealed bases
Tm  =  61°C
luciferase
859 .. 2511  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
859 .. 2511  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
AmpR
5240 .. 6100  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5240 .. 6031  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5240 .. 6100  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6032 .. 6100  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5240 .. 6100  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR
3258 .. 4052  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR
3258 .. 4052  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
4452 .. 5040  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4452 .. 5040  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
2870 .. 3227  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2870 .. 3227  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
227 .. 530  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
227 .. 530  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
531 .. 734  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
531 .. 734  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
2555 .. 2676  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2555 .. 2676  =  122 bp
SV40 polyadenylation signal
pause site
6267 .. 6358  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
6267 .. 6358  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
poly(A) signal
4077 .. 4125  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4077 .. 4125  =  49 bp
synthetic polyadenylation signal
poly(A) signal
6205 .. 6253  =  49 bp
synthetic polyadenylation signal
poly(A) signal
6205 .. 6253  =  49 bp
synthetic polyadenylation signal
SV40 ori
3078 .. 3213  =  136 bp
SV40 origin of replication
SV40 ori
3078 .. 3213  =  136 bp
SV40 origin of replication
ORF:  859 .. 2511  =  1653 bp
ORF:  550 amino acids  =  60.6 kDa
ORF:  3258 .. 4052  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5370 .. 5636  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  3437 .. 3700  =  264 bp
ORF:  87 amino acids  =  9.4 kDa
ORF:  5240 .. 6100  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  805 .. 1086  =  282 bp
ORF:  93 amino acids  =  9.9 kDa
ORF:  3372 .. 3743  =  372 bp
ORF:  123 amino acids  =  12.3 kDa
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