pGL4.78[hRlucCP Hygro]

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

Sequence Author: Promega

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AarI - BfuAI - BspMI (5273) RVprimer3 (5252 .. 5271) poly(A) signal BsmBI (5100) SpeI (5087) BstZ17I (4768) SacII (4652) PvuI (4628) BstEII (4183) BstXI (4180) AleI (4178) NotI (4156) BspHI (4056) AlwNI (3752) ApaLI (3650) BciVI (3539) PciI (3336) RVprimer4 (3137 .. 3156) SalI (3086) BstBI (3072) AgeI (3007) PmeI (2996) BssHII (2971) BtgZI (2853) 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) PfoI * (380) BsaI (640) NruI (761) ZraI (795) PflFI - Tth111I (796) AatII (797) PasI (874) EcoRI (1032) AflII (1062) hCL1 FseI (1238) AanI - PsiI (1359) HpaI (1379) MfeI (1388) BamHI (1481) PvuII (1591) SexAI * (1682) SV40 promoter StuI (1914) AvrII (1915) BsaAI (2146) PspOMI (2369) ApaI (2373) pGL4.78[hRlucCP/Hygro] 5303 bp
AarI  (5273)
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  (5273)
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  (5273)
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  (5100)
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  (5087)
1 site
A C T A G T T G A T C A
BstZ17I  (4768)
1 site
G T A T A C C A T A T G
SacII  (4652)
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  (4628)
1 site
C G A T C G G C T A G C
BstEII  (4183)
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  (4180)
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.
AleI  (4178)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (4156)
1 site
G C G G C C G C C G C C G G C G
BspHI  (4056)
1 site
T C A T G A A G T A C T
AlwNI  (3752)
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  (3650)
1 site
G T G C A C C A C G T G
BciVI  (3539)
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  (3336)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SalI  (3086)
1 site
G T C G A C C A G C T G
BstBI  (3072)
1 site
T T C G A A A A G C T T
AgeI  (3007)
1 site
A C C G G T T G G C C A
PmeI  (2996)
1 site
G T T T A A A C C A A A T T T G
BssHII  (2971)
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.
BtgZI  (2853)
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.
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.
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.
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
AflII  (1062)
1 site
C T T A A G G A A T T C
FseI  (1238)
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.
AanI  (1359)
1 site
T T A T A A A A T A T T
PsiI  (1359)
1 site
T T A T A A A A T A T T
HpaI  (1379)
1 site
G T T A A C C A A T T G
MfeI  (1388)
1 site
C A A T T G G T T A A C
BamHI  (1481)
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.
PvuII  (1591)
1 site
C A G C T G G T C G A C
SexAI  (1682)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
StuI  (1914)
1 site
A G G C C T T C C G G A
AvrII  (1915)
1 site
C C T A G G G G A T C C
BsaAI  (2146)
1 site
Y A C G T R R T G C A Y
PspOMI  (2369)
1 site
G G G C C C C C C G G G
ApaI  (2373)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
RVprimer3
20-mer  /  50% GC
1 binding site
5252 .. 5271  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
3137 .. 3156  =  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
hCL1
1039 .. 1086  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring ubiquitin-dependent degradation
human codon-optimized
hCL1
1039 .. 1086  =  48 bp
16 amino acids  =  1.9 kDa
Product: non-ORF yeast peptide conferring ubiquitin-dependent degradation
human codon-optimized
hPEST
1090 .. 1209  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
hPEST
1090 .. 1209  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
HygR
1961 .. 2998  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
1961 .. 2998  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
AmpR
4185 .. 5045  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4185 .. 4976  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4185 .. 5045  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4977 .. 5045  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4185 .. 5045  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
3397 .. 3985  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3397 .. 3985  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1573 .. 1930  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1573 .. 1930  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
1258 .. 1379  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1258 .. 1379  =  122 bp
SV40 polyadenylation signal
pause site
5212 .. 5303  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5212 .. 5303  =  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
3022 .. 3070  =  49 bp
synthetic polyadenylation signal
poly(A) signal
3022 .. 3070  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5150 .. 5198  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5150 .. 5198  =  49 bp
synthetic polyadenylation signal
SV40 ori
1781 .. 1916  =  136 bp
SV40 origin of replication
SV40 ori
1781 .. 1916  =  136 bp
SV40 origin of replication
ORF:  100 .. 1212  =  1113 bp
ORF:  370 amino acids  =  42.3 kDa
ORF:  4315 .. 4581  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1961 .. 2998  =  1038 bp
ORF:  345 amino acids  =  38.4 kDa
ORF:  825 .. 1166  =  342 bp
ORF:  113 amino acids  =  12.7 kDa
ORF:  4185 .. 5045  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2294 .. 2719  =  426 bp
ORF:  141 amino acids  =  14.3 kDa
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