pMetLuc-Mem Reporter

In vivo imaging vector for measuring promoter activity with membrane-anchored Metridia luciferase.

Sequence Author: Clontech (TaKaRa)

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AfeI (14) pause site ScaI (4197) PfoI (2990) RsrII (2731) BsrDI (2448) PflFI - Tth111I (2333) FspI (2317) BglII (27) PaeR7I - XhoI (31) Eco53kI (36) SacI (38) HindIII (40) EcoRI (47) MCS PstI (56) SalI (57) AccI (58) Acc65I (63) KpnI (67) SacII (70) PspOMI (71) ApaI (75) BamHI (78) AgeI (84) ATG PmlI (123) BclI * (431) Bpu10I (598) BsrGI (632) XbaI * (941) MfeI (1037) HpaI (1050) BtsI - BtsαI (1126) AflII (1169) DraIII (1403) SfiI (1990) BseRI (2033) StuI (2036) EagI (2121) pMetLuc-Mem Reporter 4351 bp
AfeI  (14)
1 site		 A G C G C T T C G C G A
ScaI  (4197)
1 site		 A G T A C T T C A T G A
PfoI  (2990)
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  (2731)
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  (2448)
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  (2333)
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  (2333)
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  (2317)
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
PmlI  (123)
1 site		 C A C G T G G T G C A C
BclI  (431)
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.
Bpu10I  (598)
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  (632)
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.
XbaI  (941)
1 site		 T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1037)
1 site		 C A A T T G G T T A A C
HpaI  (1050)
1 site		 G T T A A C C A A T T G
BtsI  (1126)
1 site		 G C A G T G N N C G T C A C
BtsαI  (1126)
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  (1169)
1 site		 C T T A A G G A A T T C
DraIII  (1403)
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.
SfiI  (1990)
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  (2033)
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  (2036)
1 site		 A G G C C T T C C G G A
EagI  (2121)
1 site		 C G G C C G G C C G G C
ATG
97 .. 99  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
97 .. 99  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
TfR membrane anchor
103 .. 288  =  186 bp
62 amino acids  =  6.8 kDa
Product: signal-anchor region of the transferrin receptor
TfR membrane anchor
103 .. 288  =  186 bp
62 amino acids  =  6.8 kDa
Product: signal-anchor region of the transferrin receptor
MetLuc
322 .. 927  =  606 bp
202 amino acids  =  22.0 kDa
Product: Metridia luciferase
human codon-optimized
MetLuc
322 .. 927  =  606 bp
202 amino acids  =  22.0 kDa
Product: Metridia luciferase
human codon-optimized
NeoR/KanR
2087 .. 2881  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2087 .. 2881  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
3489 .. 4077  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3489 .. 4077  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1179 .. 1634  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1179 .. 1634  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1856 .. 2052  =  197 bp
SV40 early promoter
SV40 promoter
1856 .. 2052  =  197 bp
SV40 early promoter
SV40 poly(A) signal
1051 .. 1172  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1051 .. 1172  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1661 .. 1765  =  105 bp
AmpR promoter
1661 .. 1765  =  105 bp
pause site
4201 .. 4292  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
4201 .. 4292  =  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
4139 .. 4187  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4139 .. 4187  =  49 bp
synthetic polyadenylation signal
HSV TK poly(A) signal
3113 .. 3160  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3113 .. 3160  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
1903 .. 2038  =  136 bp
SV40 origin of replication
SV40 ori
1903 .. 2038  =  136 bp
SV40 origin of replication
ORF:  97 .. 930  =  834 bp
ORF:  277 amino acids  =  30.1 kDa
ORF:  2902 .. 3351  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2087 .. 2881  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  777 .. 1028  =  252 bp
ORF:  83 amino acids  =  9.6 kDa
ORF:  2259 .. 2645  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2396 .. 2932  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3107 .. 3340  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  25 .. 960  =  936 bp
ORF:  311 amino acids  =  31.4 kDa
ORF:  4291 .. 215  =  276 bp
ORF:  91 amino acids
ORF:  279 .. 638  =  360 bp
ORF:  119 amino acids  =  12.8 kDa
ORF:  657 .. 914  =  258 bp
ORF:  85 amino acids  =  9.4 kDa
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phRG-TK
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phRL-null
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pGL4.32[luc2P NF-kB-RE Hygro]
phRL-SV40
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pCMV-CLuc 2
pGL4.33[luc2P SRE Hygro]
phRL-TK
pSF-CMV-RLuc
pCMV-Cypridina Luc
pGL4.34[luc2P SRF-RE Hygro]
phRL-TK(Int-)
pSF-CMV-RSV-FLuc AscI
pCMV-Gaussia Luc
pGL4.35[luc2P 9XGAL4UAS Hygro]
pLightSwitch_Prom
pSF-CMV-RSV-RLuc AscI
pCMV-Gaussia-Dura Luc
pGL4.36[luc2P MMTV Hygro]
pLVX-MetLuc Control
pSF-CMV-Ub-FLuc AscI
pCMV-GLuc 2
pGL4.37[luc2P ARE Hygro]
pLVX-MetLuc Reporter
pSF-CMV-Ub-RLuc AscI
pCMV-Green Renilla Luc
pGL4.38[luc2P p53 RE Hygro]
pMCS-Cypridina Luc
pSF-CMVe-FLuc
pCMV-Red Firefly Luc
pGL4.39[luc2P ATF6 RE Hygro]
pMCS-Gaussia Luc
pSF-CMVe-RLuc
pCRE-MetLuc2-Reporter
pGL4.40[luc2P MRE Hygro]
pMCS-Gaussia-Dura Luc
pSF-MinProm-FLuc
pFN26A (BIND) hRluc-neo Flexi Vector
pGL4.41[luc2P HSE Hygro]
pMCS-Green Renilla Luc
pSF-MinProm-RLuc
pGEM-luc
pGL4.42[luc2P HRE Hygro]
pMCS-Red Firefly Luc
pSF-pA-CMVe-FLuc
pGL2-Basic
pGL4.43[luc2P XRE Hygro]
pMetLuc-Mem Control
pSF-pA-CMVe-RLuc
pGL2-Control
pGL4.44[luc2P AP1 RE Hygro]
pMetLuc-Mem Reporter
pSF-pA-MinProm-FLuc
pGL2-Enhancer
pGL4.45[luc2P ISRE Hygro]
pMetLuc2-Control
pSF-pA-MinProm-RLuc
pGL2-Promoter
pGL4.47[luc2P SIE Hygro]
pMetLuc2-Reporter
pSF-pA-PromMCS-FLuc
pGL3-Basic
pGL4.48[luc2P SBE Hygro]
pmirGLO
pSF-pA-PromMCS-RLuc
pGL3-Control
pGL4.49[luc2P TCF-LEF RE Hygro]
pMirTarget
pSF-PromMCS-FLuc
pGL3-Enhancer
pGL4.50[luc2 CMV Hygro]
pNFkB-MetLuc2-Reporter
pSF-PromMCS-RLuc
pGL3-Promoter
pGL4.51[luc2 CMV Neo]
pNL1.1.CMV[Nluc CMV]
pSF-RecA-FLuc
pGL4.10[luc2]
pGL4.52[luc2P STAT5 RE Hygro]
pNL1.1[Nluc]
pSP-luc+NF
pGL4.11[luc2P]
pGL4.70[hRluc]
pNL1.2[NlucP]
pSV40-CLuc
pGL4.12[luc2CP]
pGL4.71[hRlucP]
pNL1.3.CMV[secNluc CMV]
pSV40-GLuc
pGL4.13[luc2 SV40]
pGL4.72[hRlucCP]
pNL1.3[secNluc]
pTK-CLuc
pGL4.14[luc2 Hygro]
pGL4.73[hRluc SV40]
pNL2.1[Nluc Hygro]
pTK-Cypridina Luc
pGL4.15[luc2P Hygro]
pGL4.74[hRluc TK]
pNL2.2[NlucP Hygro]
pTK-Gaussia Luc
pGL4.16[luc2CP Hygro]
pGL4.75[hRluc CMV]
pNL2.3[secNluc Hygro]
pTK-Gaussia-Dura Luc
pGL4.17[luc2 Neo]
pGL4.76[hRluc Hygro]
pNL3.1[Nluc minP]
pTK-GLuc
pGL4.18[luc2P Neo]
pGL4.77[hRlucP Hygro]
pNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]
pTK-Green Renilla Luc
pGL4.19[luc2CP Neo]
pGL4.78[hRlucCP Hygro]
pNL3.2[NlucP minP]
pTK-Red Firefly Luc
pGL4.20[luc2 Puro]
pGL4.79[hRluc Neo]
pNL3.3[secNluc minP]
pXPG
pGL4.21[luc2P Puro]
pGL4.80[hRlucP Neo]
pRL-CMV

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