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Plasmid Files

pMetLuc2-Control

Mammalian vector for constitutive expression of secreted Metridia luciferase.

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pMetLuc2-Control Sequence and MappMetLuc2-Control.dna
Map and Sequence File   
Sequence Author:  Clontech
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 ScaI (4627) PfoI (3420) BstBI (3327) RsrII (3161) BsrDI (2878) PflFI - Tth111I (2763) FspI (2747) StuI (2466) BseRI (2463) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (615) AleI (652) EcoNI (670) BclI * (788) XcmI (828) TspMI - XmaI (935) SmaI (937) Bpu10I (955) BsrGI (989) NotI (1289) XbaI * (1299) MfeI (1395) HincII - HpaI (1408) Bts α I (1484) AflII (1527) DraIII (1761) SexAI * (2234) SfiI (2420) pMetLuc2-Control 4781 bp
ScaI  (4627)
1 site
A G T A C T T C A T G A
PfoI  (3420)
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.
BstBI  (3327)
1 site
T T C G A A A A G C T T
RsrII  (3161)
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  (2878)
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  (2763)
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  (2763)
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  (2747)
1 site
T G C G C A A C G C G T
StuI  (2466)
1 site
A G G C C T T C C G G A
BseRI  (2463)
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.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
AgeI  (615)
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  (652)
1 site
C A C N N N N G T G G T G N N N N C A C
EcoNI  (670)
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  (788)
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  (828)
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.
TspMI  (935)
1 site
C C C G G G G G G C C C
XmaI  (935)
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  (937)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
Bpu10I  (955)
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  (989)
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  (1289)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1299)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1395)
1 site
C A A T T G G T T A A C
HincII  (1408)
1 site
G T Y R A C C A R Y T G
HpaI  (1408)
1 site
G T T A A C C A A T T G
BtsαI  (1484)
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  (1527)
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  (1761)
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  (2234)
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  (2420)
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.
NeoR/KanR
2517 .. 3311  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
2517 .. 3311  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
MetLuc
628 .. 1287  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 1:  signal sequence  
   628 .. 678  =  51 bp
   17 amino acids  =  1.9 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
628 .. 1287  =  660 bp
219 amino acids  =  23.9 kDa
   Segment 2:  
   679 .. 1287  =  609 bp
   202 amino acids  =  22.0 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
628 .. 1287  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
Product: secreted Metridia luciferase
human codon-optimized
ori
3919 .. 4507  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3919 .. 4507  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1537 .. 1992  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1537 .. 1992  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2125 .. 2482  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2125 .. 2482  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1409 .. 1530  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1409 .. 1530  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2019 .. 2123  =  105 bp
AmpR promoter
2019 .. 2123  =  105 bp
pause site
4631 .. 4722  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4631 .. 4722  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
poly(A) signal
4569 .. 4617  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4569 .. 4617  =  49 bp
synthetic polyadenylation signal
HSV TK poly(A) signal
3543 .. 3590  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3543 .. 3590  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2333 .. 2468  =  136 bp
SV40 origin of replication
SV40 ori
2333 .. 2468  =  136 bp
SV40 origin of replication
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