pCMV-Green Renilla Luc

Control vector for constitutive high-level expression of intracellular green Renilla luciferase under control of the CMV promoter.

Sequence Author: Thermo Fisher (Pierce)

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PaqCI (5830) EcoRV (5536) ApaI (5196) PspOMI (5192) AflII (5183) DrdI (4850) AlwNI (4543) MauBI (4254) NruI (4181) ScaI (3737) PvuI (3627) FspI (3479) AfeI (3307) SalI (3147) BstZ17I (3142) BsmI (3090) NaeI (2955) SpeI (1) NdeI (236) SnaBI (342) Eco53kI (568) SacI (570) BamHI (634) XcmI (1068) EcoNI (1321) NotI (1583) BseRI (2148) AvrII (2152) TspMI - XmaI (2173) SmaI (2175) PflFI - Tth111I (2323) BsiWI (2337) RsrII (2397) BstEII (2415) SacII (2495) BsmBI - Esp3I (2673) NgoMIV (2953) pCMV-Green Renilla Luc 5860 bp
PaqCI  (5830)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
EcoRV  (5536)
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.
ApaI  (5196)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (5192)
1 site
G G G C C C C C C G G G
AflII  (5183)
1 site
C T T A A G G A A T T C
DrdI  (4850)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AlwNI  (4543)
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.
MauBI  (4254)
1 site
C G C G C G C G G C G C G C G C
NruI  (4181)
1 site
T C G C G A A G C G C T
ScaI  (3737)
1 site
A G T A C T T C A T G A
PvuI  (3627)
1 site
C G A T C G G C T A G C
FspI  (3479)
1 site
T G C G C A A C G C G T
AfeI  (3307)
1 site
A G C G C T T C G C G A
SalI  (3147)
1 site
G T C G A C C A G C T G
BstZ17I  (3142)
1 site
G T A T A C C A T A T G
BsmI  (3090)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
NaeI  (2955)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
SpeI  (1)
1 site
A C T A G T T G A T C A
NdeI  (236)
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.
SnaBI  (342)
1 site
T A C G T A A T G C A T
Eco53kI  (568)
1 site
G A G C T C C T C G A G
SacI  (570)
1 site
G A G C T C C T C G A G
BamHI  (634)
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.
XcmI  (1068)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
EcoNI  (1321)
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.
NotI  (1583)
1 site
G C G G C C G C C G C C G G C G
BseRI  (2148)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
AvrII  (2152)
1 site
C C T A G G G G A T C C
TspMI  (2173)
1 site
C C C G G G G G G C C C
XmaI  (2173)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2175)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PflFI  (2323)
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  (2323)
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.
BsiWI  (2337)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
RsrII  (2397)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstEII  (2415)
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.
SacII  (2495)
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.
BsmBI  (2673)
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.
Esp3I  (2673)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
NgoMIV  (2953)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
Green Renilla Luciferase
646 .. 1581  =  936 bp
311 amino acids  =  36.0 kDa
Product: green-emitting variant of Renilla luciferase
human codon-optimized
Green Renilla Luciferase
646 .. 1581  =  936 bp
311 amino acids  =  36.0 kDa
Product: green-emitting variant of Renilla luciferase
human codon-optimized
AmpR
3184 .. 4044  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3184 .. 3975  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3184 .. 4044  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3976 .. 4044  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3184 .. 4044  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
2281 .. 2880  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2281 .. 2880  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4308 .. 4896  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4308 .. 4896  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1838 .. 2167  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1838 .. 2167  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
63 .. 366  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
63 .. 366  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
367 .. 570  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
367 .. 570  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
3010 .. 3131  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3010 .. 3131  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
1604 .. 1715  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1604 .. 1715  =  112 bp
bovine growth hormone polyadenylation signal
pause site
5769 .. 5860  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5769 .. 5860  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
EM7 promoter
2215 .. 2262  =  48 bp
synthetic bacterial promoter
EM7 promoter
2215 .. 2262  =  48 bp
synthetic bacterial promoter
T7 promoter
615 .. 633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
615 .. 633  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
5638 .. 5654  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
5638 .. 5654  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
SV40 ori
2018 .. 2153  =  136 bp
SV40 origin of replication
SV40 ori
2018 .. 2153  =  136 bp
SV40 origin of replication
ORF:  526 .. 1581  =  1056 bp
ORF:  351 amino acids  =  40.8 kDa
ORF:  2281 .. 2880  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  3314 .. 3580  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2441 .. 2767  =  327 bp
ORF:  108 amino acids  =  12.0 kDa
ORF:  3184 .. 4044  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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