pGL4.80[hRlucP Neo]

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

Sequence Author: Promega

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AarI (4980) RVprimer3 (4959 .. 4978) poly(A) signal BsmBI (4807) SpeI (4794) BstZ17I (4475) SacII (4359) PvuI (4335) AhdI (3965) BstEII (3890) BstXI - PstI (3887) AleI (3885) NotI (3863) BspHI (3763) ApaLI (3357) BciVI (3246) BssSI (3216) AflIII - PciI (3043) RVprimer4 (2844 .. 2863) SalI (2793) BstBI (2779) AgeI (2714) BbvCI (2595) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) SacI (25) NheI (27) BmtI (31) AbsI - PaeR7I - PspXI - XhoI (33) BglII (46) BglI - SfiI (59) HindIII (65) BglI (100) DraIII (288) PfoI * (380) XmnI (634) BsaI (640) NruI (761) ZraI (795) PflFI - Tth111I (796) AatII (797) PasI (874) EcoRI (1032) AanI - PsiI (1308) HpaI (1328) MfeI (1337) BamHI (1430) FspI (1468) PvuII (1540) SV40 promoter StuI (1863) PspOMI (1980) ApaI (1984) SgrAI (2126) EcoNI (2384) pGL4.80[hRlucP/Neo] 5010 bp
AarI  (4980)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BsmBI  (4807)
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.
BsmBI-v2 is an improved version of BsmBI.
SpeI  (4794)
1 site
A C T A G T T G A T C A
BstZ17I  (4475)
1 site
G T A T A C C A T A T G
SacII  (4359)
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  (4335)
1 site
C G A T C G G C T A G C
AhdI  (3965)
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  (3890)
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  (3887)
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  (3887)
1 site
C T G C A G G A C G T C
AleI  (3885)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (3863)
1 site
G C G G C C G C C G C C G G C G
BspHI  (3763)
1 site
T C A T G A A G T A C T
ApaLI  (3357)
1 site
G T G C A C C A C G T G
BciVI  (3246)
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.
BssSI  (3216)
1 site
C A C G A G G T G C T C
AflIII  (3043)
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  (3043)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (2793)
1 site
G T C G A C C A G C T G
BstBI  (2779)
1 site
T T C G A A A A G C T T
AgeI  (2714)
1 site
A C C G G T T G G C C A
BbvCI  (2595)
1 site
C C T C A G C G G A G T C G
BglI  (8)
3 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
BglII  (46)
1 site
A G A T C T T C T A G A
BglI  (59)
3 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
BglI  (100)
3 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.
DraIII  (288)
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  (380)
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.
XmnI  (634)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaI  (640)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
NruI  (761)
1 site
T C G C G A A G C G C T
ZraI  (795)
1 site
G A C G T C C T G C A G
PflFI  (796)
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  (796)
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  (797)
1 site
G A C G T C C T G C A G
PasI  (874)
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.
EcoRI  (1032)
1 site
G A A T T C C T T A A G
AanI  (1308)
1 site
T T A T A A A A T A T T
PsiI  (1308)
1 site
T T A T A A A A T A T T
HpaI  (1328)
1 site
G T T A A C C A A T T G
MfeI  (1337)
1 site
C A A T T G G T T A A C
BamHI  (1430)
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.
FspI  (1468)
1 site
T G C G C A A C G C G T
PvuII  (1540)
1 site
C A G C T G G T C G A C
StuI  (1863)
1 site
A G G C C T T C C G G A
PspOMI  (1980)
1 site
G G G C C C C C C G G G
ApaI  (1984)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SgrAI  (2126)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
EcoNI  (2384)
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.
RVprimer3
20-mer  /  50% GC
1 binding site
4959 .. 4978  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
2844 .. 2863  =  20 annealed bases
Tm  =  62°C
hRluc
100 .. 1032  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
100 .. 1032  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hPEST
1036 .. 1155  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
hPEST
1036 .. 1155  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
AmpR
3892 .. 4752  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3892 .. 4683  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3892 .. 4752  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4684 .. 4752  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3892 .. 4752  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR
1910 .. 2704  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR
1910 .. 2704  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3104 .. 3692  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3104 .. 3692  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1522 .. 1879  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1522 .. 1879  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
1207 .. 1328  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1207 .. 1328  =  122 bp
SV40 polyadenylation signal
pause site
4919 .. 5010  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
4919 .. 5010  =  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
2729 .. 2777  =  49 bp
synthetic polyadenylation signal
poly(A) signal
2729 .. 2777  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4857 .. 4905  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4857 .. 4905  =  49 bp
synthetic polyadenylation signal
SV40 ori
1730 .. 1865  =  136 bp
SV40 origin of replication
SV40 ori
1730 .. 1865  =  136 bp
SV40 origin of replication
ORF:  100 .. 1158  =  1059 bp
ORF:  352 amino acids  =  40.2 kDa
ORF:  1910 .. 2704  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  4022 .. 4288  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2089 .. 2352  =  264 bp
ORF:  87 amino acids  =  9.4 kDa
ORF:  3892 .. 4752  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2024 .. 2395  =  372 bp
ORF:  123 amino acids  =  12.3 kDa
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Download pGL4.80[hRlucP Neo].dna file

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