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Plasmid Files

pCMV-Green Renilla Luc

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

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pCMV-Green Renilla Luc Sequence and MappCMV-Green Renilla Luc.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (Pierce)
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 AarI (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 (2673) NgoMIV (2953) pCMV-Green Renilla Luc 5860 bp
AarI  (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 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.
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

The sticky ends produced by AflII are hard to ligate.
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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.
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
   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
   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
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