pGL4.34[luc2P SRF-RE Hygro]

Vector with a minimal promoter and a serum response element for studying cell signaling using destabilized luciferase.

Sequence Author: Promega

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AarI - BfuAI - BspMI (6082) RVprimer3 (6061 .. 6080) BsmBI (5909) SpeI (5896) BstZ17I (5577) SacII (5461) PvuI (5437) Bsu36I (5423) BstEII (4992) BstXI - PstI (4989) NotI (4965) AlwNI (4561) ApaLI (4459) BciVI (4348) PciI (4145) RVprimer4 (3946 .. 3965) SalI (3895) BstBI (3881) PmeI (3805) BssHII (3780) BsaAI (2955) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) SacI (25) NheI (27) BmtI (31) BglII (148) BglI - SfiI (161) HindIII (167) MreI - SgrAI (306) BsrGI (733) BbvCI (954) KasI (1190) NarI (1191) SfoI (1192) PluTI (1194) BlpI (1197) DraIII (1389) BpmI (1623) EcoRI (1892) FseI (2047) PsiI (2168) MfeI (2197) BamHI (2290) StuI (2723) AvrII (2724) XmnI (2807) pGL4.34[luc2P/SRF-RE/Hygro] 6112 bp
AarI  (6082)
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.
BfuAI  (6082)
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  (6082)
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.
BsmBI  (5909)
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  (5896)
1 site
A C T A G T T G A T C A
BstZ17I  (5577)
1 site
G T A T A C C A T A T G
SacII  (5461)
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  (5437)
1 site
C G A T C G G C T A G C
Bsu36I  (5423)
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.
BstEII  (4992)
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  (4989)
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  (4989)
1 site
C T G C A G G A C G T C
NotI  (4965)
1 site
G C G G C C G C C G C C G G C G
AlwNI  (4561)
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.
ApaLI  (4459)
1 site
G T G C A C C A C G T G
BciVI  (4348)
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.
PciI  (4145)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (3895)
1 site
G T C G A C C A G C T G
BstBI  (3881)
1 site
T T C G A A A A G C T T
PmeI  (3805)
1 site
G T T T A A A C C A A A T T T G
BssHII  (3780)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BsaAI  (2955)
1 site
Y A C G T R R T G C A Y
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
BglII  (148)
1 site
A G A T C T T C T A G A
BglI  (161)
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  (161)
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  (167)
1 site
A A G C T T T T C G A A
MreI  (306)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (306)
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.
BsrGI  (733)
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.
BbvCI  (954)
1 site
C C T C A G C G G A G T C G
KasI  (1190)
1 site
G G C G C C C C G C G G
NarI  (1191)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1192)
1 site
G G C G C C C C G C G G
PluTI  (1194)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BlpI  (1197)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
DraIII  (1389)
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  (1623)
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  (1892)
1 site
G A A T T C C T T A A G
FseI  (2047)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
PsiI  (2168)
1 site
T T A T A A A A T A T T
MfeI  (2197)
1 site
C A A T T G G T T A A C
BamHI  (2290)
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  (2723)
1 site
A G G C C T T C C G G A
AvrII  (2724)
1 site
C C T A G G G G A T C C
XmnI  (2807)
1 site
G A A N N N N T T C C T T N N N N A A G
RVprimer3
20-mer  /  50% GC
1 binding site
6061 .. 6080  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
3946 .. 3965  =  20 annealed bases
Tm  =  62°C
luciferase
243 .. 1892  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
luciferase
243 .. 1892  =  1650 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luc2 version of the luciferase gene
hPEST
1896 .. 2015  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
hPEST
1896 .. 2015  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
HygR
2770 .. 3807  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
2770 .. 3807  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
AmpR
4994 .. 5854  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4994 .. 5785  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4994 .. 5854  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5786 .. 5854  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4994 .. 5854  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
4206 .. 4794  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4206 .. 4794  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
2382 .. 2739  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2382 .. 2739  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2067 .. 2188  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2067 .. 2188  =  122 bp
SV40 polyadenylation signal
SRF-RE
38 .. 143  =  106 bp
serum response factor response element (5 copies)
SRF-RE
38 .. 143  =  106 bp
serum response factor response element (5 copies)
pause site
6021 .. 6112  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
6021 .. 6112  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
poly(A) signal
3831 .. 3879  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3831 .. 3879  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5959 .. 6007  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5959 .. 6007  =  49 bp
synthetic polyadenylation signal
minP
179 .. 210  =  32 bp
minimal TATA-box promoter with low basal activity
minP
179 .. 210  =  32 bp
minimal TATA-box promoter with low basal activity
SV40 ori
2590 .. 2725  =  136 bp
SV40 origin of replication
SV40 ori
2590 .. 2725  =  136 bp
SV40 origin of replication
ORF:  2770 .. 3807  =  1038 bp
ORF:  345 amino acids  =  38.4 kDa
ORF:  192 .. 2018  =  1827 bp
ORF:  608 amino acids  =  66.7 kDa
ORF:  5124 .. 5390  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4994 .. 5854  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3103 .. 3528  =  426 bp
ORF:  141 amino acids  =  14.3 kDa
ORF:  171 .. 470  =  300 bp
ORF:  99 amino acids  =  10.7 kDa
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