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Plasmid Files

pCMV-Red Firefly Luc

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

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pCMV-Red Firefly Luc Sequence and MappCMV-Red Firefly Luc.dna
Map and Sequence File   
Sequence Author:  Thermo Scientific (Pierce)
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 AarI - BfuAI - BspMI (6545) EcoRV (6251) AflII (5898) DrdI (5565) MauBI (4969) NruI (4896) ScaI (4452) PvuI (4342) FspI (4194) AfeI (4022) SalI (3862) BstZ17I (3857) BsmI (3805) DraIII (3567) BsmBI (3388) SpeI (2) CMV enhancer NdeI (237) SnaBI (343) Eco53kI (569) SacI (571) BamHI (635) BsrDI (768) EcoRI (911) BglII (972) PflMI * (1141) KflI - PpuMI (1321) SbfI (1485) MreI (2051) NotI (2298) BseRI (2863) AvrII (2867) TspMI - XmaI (2888) SmaI (2890) BsiWI (3052) RsrII (3112) BstEII (3130) SacII (3210) pCMV-Red Firefly Luc 6575 bp
AarI  (6545)
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.
BfuAI  (6545)
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  (6545)
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.
EcoRV  (6251)
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.
AflII  (5898)
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  (5565)
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.
MauBI  (4969)
1 site
C G C G C G C G G C G C G C G C
NruI  (4896)
1 site
T C G C G A A G C G C T
ScaI  (4452)
1 site
A G T A C T T C A T G A
PvuI  (4342)
1 site
C G A T C G G C T A G C
FspI  (4194)
1 site
T G C G C A A C G C G T
AfeI  (4022)
1 site
A G C G C T T C G C G A
SalI  (3862)
1 site
G T C G A C C A G C T G
BstZ17I  (3857)
1 site
G T A T A C C A T A T G
BsmI  (3805)
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.
DraIII  (3567)
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.
BsmBI  (3388)
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.
SpeI  (2)
1 site
A C T A G T T G A T C A
NdeI  (237)
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  (343)
1 site
T A C G T A A T G C A T
Eco53kI  (569)
1 site
G A G C T C C T C G A G
SacI  (571)
1 site
G A G C T C C T C G A G
BamHI  (635)
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.
BsrDI  (768)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
EcoRI  (911)
1 site
G A A T T C C T T A A G
BglII  (972)
1 site
A G A T C T T C T A G A
PflMI  (1141)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
KflI  (1321)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1321)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
SbfI  (1485)
1 site
C C T G C A G G G G A C G T C C
MreI  (2051)
1 site
C G C C G G C G G C G G C C G C
NotI  (2298)
1 site
G C G G C C G C C G C C G G C G
BseRI  (2863)
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  (2867)
1 site
C C T A G G G G A T C C
TspMI  (2888)
1 site
C C C G G G G G G C C C
XmaI  (2888)
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  (2890)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BsiWI  (3052)
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  (3112)
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  (3130)
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  (3210)
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.
Red Firefly Luciferase
647 .. 2293  =  1647 bp
548 amino acids  =  60.1 kDa
Product: intracellular red firefly luciferase
human codon-optimized
Red Firefly Luciferase
647 .. 2293  =  1647 bp
548 amino acids  =  60.1 kDa
Product: intracellular red firefly luciferase
human codon-optimized
AmpR
3899 .. 4759  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3899 .. 4690  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3899 .. 4759  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4691 .. 4759  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3899 .. 4759  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
2996 .. 3595  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2996 .. 3595  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
5023 .. 5611  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5023 .. 5611  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 promoter
2553 .. 2882  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
2553 .. 2882  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
64 .. 367  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
64 .. 367  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
368 .. 571  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
368 .. 571  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
3725 .. 3846  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3725 .. 3846  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
2319 .. 2430  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
2319 .. 2430  =  112 bp
bovine growth hormone polyadenylation signal
pause site
6484 .. 6575  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
6484 .. 6575  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
EM7 promoter
2930 .. 2977  =  48 bp
synthetic bacterial promoter
EM7 promoter
2930 .. 2977  =  48 bp
synthetic bacterial promoter
T7 promoter
616 .. 634  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
616 .. 634  =  19 bp
promoter for bacteriophage T7 RNA polymerase
lac operator
6353 .. 6369  =  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
6353 .. 6369  =  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
2733 .. 2868  =  136 bp
SV40 origin of replication
SV40 ori
2733 .. 2868  =  136 bp
SV40 origin of replication
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