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Plasmid Files

pMetLuc2-Reporter

Mammalian vector for measuring the activity of promoter and enhancer sequences using secreted Metridia luciferase.

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pMetLuc2-Reporter Sequence and MappMetLuc2-Reporter.dna
Map and Sequence File   
Sequence Author:  Clontech
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 AfeI (14) ScaI (4096) poly(A) signal PfoI (2889) RsrII (2630) BsrDI (2347) PflFI - Tth111I (2232) FspI (2216) BglII (27) PaeR7I - XhoI (31) Eco53kI (36) SacI (38) HindIII (40) EcoRI (47) PstI (56) SalI (57) AccI (58) Acc65I (63) KpnI (67) SacII (70) PspOMI (71) ApaI (75) BamHI (78) AgeI (84) AleI (121) EcoNI (139) BclI * (257) XcmI (297) Bpu10I (424) BsrGI (458) NotI (758) XbaI * (768) MfeI (864) HpaI (877) Bts α I (953) AflII (996) DraIII (1230) SexAI * (1703) SV40 promoter SfiI (1889) BseRI (1932) StuI (1935) pMetLuc2-Reporter 4250 bp
AfeI  (14)
1 site
A G C G C T T C G C G A
ScaI  (4096)
1 site
A G T A C T T C A T G A
PfoI  (2889)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
RsrII  (2630)
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.
BsrDI  (2347)
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.
PflFI  (2232)
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  (2232)
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.
FspI  (2216)
1 site
T G C G C A A C G C G T
BglII  (27)
1 site
A G A T C T T C T A G A
PaeR7I  (31)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (31)
1 site
C T C G A G G A G C T C
Eco53kI  (36)
1 site
G A G C T C C T C G A G
SacI  (38)
1 site
G A G C T C C T C G A G
HindIII  (40)
1 site
A A G C T T T T C G A A
EcoRI  (47)
1 site
G A A T T C C T T A A G
PstI  (56)
1 site
C T G C A G G A C G T C
SalI  (57)
1 site
G T C G A C C A G C T G
AccI  (58)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
Acc65I  (63)
1 site
G G T A C C C C A T G G
KpnI  (67)
1 site
G G T A C C C C A T G G
SacII  (70)
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.
PspOMI  (71)
1 site
G G G C C C C C C G G G
ApaI  (75)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BamHI  (78)
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.
AgeI  (84)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AleI  (121)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoNI  (139)
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.
BclI  (257)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
XcmI  (297)
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.
Bpu10I  (424)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BsrGI  (458)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
NotI  (758)
1 site
G C G G C C G C C G C C G G C G
XbaI  (768)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (864)
1 site
C A A T T G G T T A A C
HpaI  (877)
1 site
G T T A A C C A A T T G
BtsαI  (953)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
AflII  (996)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
DraIII  (1230)
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.
SexAI  (1703)
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.
SfiI  (1889)
1 site
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.
BseRI  (1932)
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.
StuI  (1935)
1 site
A G G C C T T C C G G A
NeoR/KanR
1986 .. 2780  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
1986 .. 2780  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
MetLuc
97 .. 756  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 1:  signal sequence  
   97 .. 147  =  51 bp
   17 amino acids  =  1.9 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
97 .. 756  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 2:  
   148 .. 756  =  609 bp
   202 amino acids  =  22.0 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
97 .. 756  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
Product: secreted Metridia luciferase
human codon-optimized
ori
3388 .. 3976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3388 .. 3976  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1006 .. 1461  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1006 .. 1461  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1594 .. 1951  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1594 .. 1951  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
878 .. 999  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
878 .. 999  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1488 .. 1592  =  105 bp
AmpR promoter
1488 .. 1592  =  105 bp
pause site
4100 .. 4191  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4100 .. 4191  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
MCS
12 .. 89  =  78 bp
multiple cloning site
MCS
12 .. 89  =  78 bp
multiple cloning site
poly(A) signal
4038 .. 4086  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4038 .. 4086  =  49 bp
synthetic polyadenylation signal
HSV TK poly(A) signal
3012 .. 3059  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3012 .. 3059  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
1802 .. 1937  =  136 bp
SV40 origin of replication
SV40 ori
1802 .. 1937  =  136 bp
SV40 origin of replication
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