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Plasmid Files

pMCS-Red Firefly Luc

Intracellular red firefly luciferase vector for cloning a transcriptional regulatory element.

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pMCS-Red Firefly Luc.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Pierce)
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Acc65I (13) SpeI (1) AarI - BfuAI - BspMI (5947) EcoRV (5653) AflII (5300) DrdI (4967) MauBI (4371) NruI (4298) ScaI (3854) PvuI (3744) FspI (3596) AfeI (3424) SalI (3264) BstZ17I (3259) BsmI (3207) KpnI (17) Eco53kI (21) SacI (23) PaeR7I - XhoI (25) HindIII (31) BamHI (37) BsrDI (170) BglII (374) PflMI * (543) KflI - PpuMI (723) SbfI (887) MreI (1453) NotI (1700) AvrII (2269) TspMI - XmaI (2290) SmaI (2292) BsiWI (2454) RsrII (2514) BstEII (2532) SacII (2612) BsmBI (2790) DraIII (2969) pMCS-Red Firefly Luc 5977 bp
Acc65I  (13)
1 site
G G T A C C C C A T G G
SpeI  (1)
1 site
A C T A G T T G A T C A
AarI  (5947)
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  (5947)
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  (5947)
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  (5653)
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  (5300)
1 site
C T T A A G G A A T T C
DrdI  (4967)
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  (4371)
1 site
C G C G C G C G G C G C G C G C
NruI  (4298)
1 site
T C G C G A A G C G C T
ScaI  (3854)
1 site
A G T A C T T C A T G A
PvuI  (3744)
1 site
C G A T C G G C T A G C
FspI  (3596)
1 site
T G C G C A A C G C G T
AfeI  (3424)
1 site
A G C G C T T C G C G A
SalI  (3264)
1 site
G T C G A C C A G C T G
BstZ17I  (3259)
1 site
G T A T A C C A T A T G
BsmI  (3207)
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.
KpnI  (17)
1 site
G G T A C C C C A T G G
Eco53kI  (21)
1 site
G A G C T C C T C G A G
SacI  (23)
1 site
G A G C T C C T C G A G
PaeR7I  (25)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (25)
1 site
C T C G A G G A G C T C
HindIII  (31)
1 site
A A G C T T T T C G A A
BamHI  (37)
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  (170)
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.
BglII  (374)
1 site
A G A T C T T C T A G A
PflMI  (543)
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  (723)
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  (723)
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  (887)
1 site
C C T G C A G G G G A C G T C C
MreI  (1453)
1 site
C G C C G G C G G C G G C C G C
NotI  (1700)
1 site
G C G G C C G C C G C C G G C G
AvrII  (2269)
1 site
C C T A G G G G A T C C
TspMI  (2290)
1 site
C C C G G G G G G C C C
XmaI  (2290)
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  (2292)
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  (2454)
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  (2514)
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  (2532)
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  (2612)
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  (2790)
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.
DraIII  (2969)
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.
Red Firefly Luciferase
49 .. 1695  =  1647 bp
548 amino acids  =  60.1 kDa
Product: intracellular red firefly luciferase
human codon-optimized
Red Firefly Luciferase
49 .. 1695  =  1647 bp
548 amino acids  =  60.1 kDa
Product: intracellular red firefly luciferase
human codon-optimized
AmpR
3301 .. 4161  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3301 .. 4092  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3301 .. 4161  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4093 .. 4161  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3301 .. 4161  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
2398 .. 2997  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2398 .. 2997  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
4425 .. 5013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4425 .. 5013  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1955 .. 2284  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
1955 .. 2284  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
3127 .. 3248  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3127 .. 3248  =  122 bp
SV40 polyadenylation signal
bGH poly(A) signal
1721 .. 1832  =  112 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1721 .. 1832  =  112 bp
bovine growth hormone polyadenylation signal
pause site
5886 .. 5977  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
5886 .. 5977  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
EM7 promoter
2332 .. 2379  =  48 bp
synthetic bacterial promoter
EM7 promoter
2332 .. 2379  =  48 bp
synthetic bacterial promoter
MCS
1 .. 42  =  42 bp
multiple cloning site
MCS
1 .. 42  =  42 bp
multiple cloning site
lac operator
5755 .. 5771  =  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
5755 .. 5771  =  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
2135 .. 2270  =  136 bp
SV40 origin of replication
SV40 ori
2135 .. 2270  =  136 bp
SV40 origin of replication
ORF:  49 .. 1695  =  1647 bp
ORF:  548 amino acids  =  60.1 kDa
ORF:  2398 .. 2997  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  3431 .. 3697  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  2558 .. 2884  =  327 bp
ORF:  108 amino acids  =  12.0 kDa
ORF:  3301 .. 4161  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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