pMetLuc-Mem Control

In vivo imaging vector for the constitutive expression of membrane-anchored Metridia luciferase.

Sequence Author: Clontech (TaKaRa)

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AflIII - PciI (4706) PfoI (3563) BstBI (3470) RsrII (3304) BsrDI (3021) PflFI - Tth111I (2906) FspI (2890) BspDI * - ClaI * (2628) StuI (2609) BseRI (2606) SfiI (2563) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) PmlI (623) BclI * (931) XcmI (971) TspMI - XmaI (1078) SmaI (1080) Bpu10I (1098) BsrGI (1132) PaqCI (1135) NotI (1432) XbaI * (1442) MfeI (1538) HincII - HpaI (1551) BtsI - BtsαI (1627) AflII (1670) DraIII (1904) CsiI - SexAI * (2377) pMetLuc-Mem Control 4764 bp
AflIII  (4706)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4706)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PfoI  (3563)
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  (3470)
1 site
T T C G A A A A G C T T
RsrII  (3304)
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  (3021)
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  (2906)
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  (2906)
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  (2890)
1 site
T G C G C A A C G C G T
BspDI  (2628)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2628)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2609)
1 site
A G G C C T T C C G G A
BseRI  (2606)
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.
SfiI  (2563)
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.
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
PmlI  (623)
1 site
C A C G T G G T G C A C
BclI  (931)
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  (971)
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  (1078)
1 site
C C C G G G G G G C C C
XmaI  (1078)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (1080)
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  (1098)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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  (1132)
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.
PaqCI  (1135)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
NotI  (1432)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1442)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1538)
1 site
C A A T T G G T T A A C
HincII  (1551)
1 site
G T Y R A C C A R Y T G
HpaI  (1551)
1 site
G T T A A C C A A T T G
BtsI  (1627)
1 site
G C A G T G N N C G T C A C
BtsαI  (1627)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
AflII  (1670)
1 site
C T T A A G G A A T T C
DraIII  (1904)
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.
CsiI  (2377)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2377)
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.
ATG
597 .. 599  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
597 .. 599  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
TfR membrane anchor
603 .. 788  =  186 bp
62 amino acids  =  6.8 kDa
Product: signal-anchor region of the transferrin receptor
TfR membrane anchor
603 .. 788  =  186 bp
62 amino acids  =  6.8 kDa
Product: signal-anchor region of the transferrin receptor
MetLuc
822 .. 1427  =  606 bp
202 amino acids  =  22.0 kDa
Product: Metridia luciferase
human codon-optimized
MetLuc
822 .. 1427  =  606 bp
202 amino acids  =  22.0 kDa
Product: Metridia luciferase
human codon-optimized
NeoR/KanR
2660 .. 3454  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2660 .. 3454  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
4062 .. 4650  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4062 .. 4650  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1680 .. 2135  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1680 .. 2135  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2268 .. 2625  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2268 .. 2625  =  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
1552 .. 1673  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1552 .. 1673  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2162 .. 2266  =  105 bp
AmpR promoter
2162 .. 2266  =  105 bp
HSV TK poly(A) signal
3686 .. 3733  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3686 .. 3733  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2476 .. 2611  =  136 bp
SV40 origin of replication
SV40 ori
2476 .. 2611  =  136 bp
SV40 origin of replication
ORF:  3475 .. 3924  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2660 .. 3454  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  597 .. 1430  =  834 bp
ORF:  277 amino acids  =  30.1 kDa
ORF:  2832 .. 3218  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  779 .. 1138  =  360 bp
ORF:  119 amino acids  =  12.8 kDa
ORF:  1157 .. 1414  =  258 bp
ORF:  85 amino acids  =  9.4 kDa
ORF:  2969 .. 3505  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3680 .. 3913  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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