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Plasmid Files

pGL4.19[luc2CP/Neo]

Promoterless vector encoding highly destabilized luciferase for measuring the activity of promoter and enhancer sequences.

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pGL4.19[luc2CP Neo] Sequence and MappGL4.19[luc2CP Neo].dna
Map and Sequence File   
Sequence Author:  Promega
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 AarI (5748) RVprimer3 (5727 .. 5746) poly(A) signal BsmBI (5575) SpeI (5562) BstZ17I (5243) SacII (5127) PvuI (5103) Bsu36I (5089) AhdI (4733) BstEII (4658) BstXI (4655) NotI (4631) ApaLI (4125) BciVI (4014) AflIII - PciI (3811) RVprimer4 (3612 .. 3631) SalI (3561) BstBI (3547) EcoNI (3152) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) MCS EcoRV (41) BglII (46) BglI - SfiI (59) HindIII (65) MreI (163) BsrGI (590) BpmI (1480) EcoRI (1749) XmnI (1768) AflII (1779) hCL1 hPEST PsiI (2076) MfeI (2105) BamHI (2198) StuI (2631) pGL4.19[luc2CP/Neo] 5778 bp
AarI  (5748)
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.
BsmBI  (5575)
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.
SpeI  (5562)
1 site
A C T A G T T G A T C A
BstZ17I  (5243)
1 site
G T A T A C C A T A T G
SacII  (5127)
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  (5103)
1 site
C G A T C G G C T A G C
Bsu36I  (5089)
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  (4733)
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  (4658)
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  (4655)
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.
NotI  (4631)
1 site
G C G G C C G C C G C C G G C G
ApaLI  (4125)
1 site
G T G C A C C A C G T G
BciVI  (4014)
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  (3811)
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  (3811)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (3561)
1 site
G T C G A C C A G C T G
BstBI  (3547)
1 site
T T C G A A A A G C T T
EcoNI  (3152)
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.
BglI  (8)
2 sites
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
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.
Acc65I  (14)
1 site
G G T A C C C C A T G G
KpnI  (18)
1 site
G G T A C C C C A T G G
Eco53kI  (23)
1 site
G A G C T C C T C G A G
SacI  (25)
1 site
G A G C T C C T C G A G
NheI  (27)
1 site
G C T A G C C G A T C G
BmtI  (31)
1 site
G C T A G C C G A T C G
AbsI  (33)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (33)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (33)
1 site
V C T C G A G B B G A G C T C V
XhoI  (33)
1 site
C T C G A G G A G C T C
EcoRV  (41)
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.
BglII  (46)
1 site
A G A T C T T C T A G A
BglI  (59)
2 sites
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SfiI  (59)
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.
HindIII  (65)
1 site
A A G C T T T T C G A A
MreI  (163)
1 site
C G C C G G C G G C G G C C G C
BsrGI  (590)
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.
BpmI  (1480)
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.
EcoRI  (1749)
1 site
G A A T T C C T T A A G
XmnI  (1768)
1 site
G A A N N N N T T C C T T N N N N A A G
AflII  (1779)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PsiI  (2076)
1 site
T T A T A A A A T A T T
MfeI  (2105)
1 site
C A A T T G G T T A A C
BamHI  (2198)
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.
StuI  (2631)
1 site
A G G C C T T C C G G A
RVprimer3
20-mer  /  50% GC
1 binding site
5727 .. 5746  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
3612 .. 3631  =  20 annealed bases
Tm  =  61°C
luciferase
100 .. 1749  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
100 .. 1749  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
AmpR
4660 .. 5520  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4660 .. 5451  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4660 .. 5520  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5452 .. 5520  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4660 .. 5520  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR
2678 .. 3472  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR
2678 .. 3472  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
3872 .. 4460  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3872 .. 4460  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 promoter
2290 .. 2647  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2290 .. 2647  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
1975 .. 2096  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1975 .. 2096  =  122 bp
SV40 polyadenylation signal
hPEST
1807 .. 1926  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse
ornithine decarboxylase
human codon-optimized
hPEST
1807 .. 1926  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse
ornithine decarboxylase
human codon-optimized
pause site
5687 .. 5778  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
5687 .. 5778  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
MCS
1 .. 70  =  70 bp
multiple cloning site
MCS
1 .. 70  =  70 bp
multiple cloning site
poly(A) signal
3497 .. 3545  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3497 .. 3545  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5625 .. 5673  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5625 .. 5673  =  49 bp
synthetic polyadenylation signal
hCL1
1756 .. 1803  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring
ubiquitin-dependent degradation
human codon-optimized
hCL1
1756 .. 1803  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring
ubiquitin-dependent degradation
human codon-optimized
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