pNFkB-MetLuc2-Reporter

Vector for monitoring NF-κB-mediated signal transduction using secreted Metridia luciferase.

Sequence Author: Clontech (TaKaRa)

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ScaI (4212) poly(A) signal RsrII (2746) BsrDI (2463) PflFI - Tth111I (2348) FspI (2332) AfeI (14) PaeR7I - XhoI (31) Eco53kI (36) SacI (38) minP PstI (172) SalI (173) AccI (174) Acc65I (179) KpnI (183) SacII (186) PspOMI (187) ApaI (191) BamHI (194) AgeI (200) AleI (237) EcoNI (255) BclI * (373) XcmI (413) Bpu10I (540) BsrGI (574) NotI (874) MfeI (980) HpaI (993) BtsI - BtsαI (1069) AflII (1112) DraIII (1346) CsiI - SexAI * (1819) SV40 promoter SfiI (2005) BseRI (2048) StuI (2051) pNF κ B-MetLuc2-Reporter 4366 bp
ScaI  (4212)
1 site		 A G T A C T T C A T G A
RsrII  (2746)
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  (2463)
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  (2348)
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  (2348)
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  (2332)
1 site		 T G C G C A A C G C G T
AfeI  (14)
1 site		 A G C G C T T C G C 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
PstI  (172)
1 site		 C T G C A G G A C G T C
SalI  (173)
1 site		 G T C G A C C A G C T G
AccI  (174)
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  (179)
1 site		 G G T A C C C C A T G G
KpnI  (183)
1 site		 G G T A C C C C A T G G
SacII  (186)
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  (187)
1 site		 G G G C C C C C C G G G
ApaI  (191)
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  (194)
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  (200)
1 site		 A C C G G T T G G C C A
AleI  (237)
1 site		 C A C N N N N G T G G T G N N N N C A C
EcoNI  (255)
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  (373)
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  (413)
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  (540)
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  (574)
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  (874)
1 site		 G C G G C C G C C G C C G G C G
MfeI  (980)
1 site		 C A A T T G G T T A A C
HpaI  (993)
1 site		 G T T A A C C A A T T G
BtsI  (1069)
1 site		 G C A G T G N N C G T C A C
BtsαI  (1069)
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  (1112)
1 site		 C T T A A G G A A T T C
DraIII  (1346)
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  (1819)
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  (1819)
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  (2005)
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  (2048)
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  (2051)
1 site		 A G G C C T T C C G G A
NeoR/KanR
2102 .. 2896  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2102 .. 2896  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
MetLuc
213 .. 872  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
   Segment 1:  signal sequence  
   213 .. 263  =  51 bp
   17 amino acids  =  1.9 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
213 .. 872  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
   Segment 2:  
   264 .. 872  =  609 bp
   202 amino acids  =  22.0 kDa
Product: secreted Metridia luciferase
human codon-optimized
MetLuc
213 .. 872  =  660 bp
219 amino acids  =  23.9 kDa
2 segments
Product: secreted Metridia luciferase
human codon-optimized
ori
3504 .. 4092  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3504 .. 4092  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1122 .. 1577  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1122 .. 1577  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1710 .. 2067  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1710 .. 2067  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
994 .. 1115  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
994 .. 1115  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1604 .. 1708  =  105 bp
AmpR promoter
1604 .. 1708  =  105 bp
pause site
4216 .. 4307  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
4216 .. 4307  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
poly(A) signal
4154 .. 4202  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4154 .. 4202  =  49 bp
synthetic polyadenylation signal
HSV TK poly(A) signal
3128 .. 3175  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3128 .. 3175  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
NF-κB RE
46 .. 85  =  40 bp
NF-κB response element (4 copies)
NF-κB RE
46 .. 85  =  40 bp
NF-κB response element (4 copies)
minP
98 .. 129  =  32 bp
minimal TATA-box promoter with low basal activity
minP
98 .. 129  =  32 bp
minimal TATA-box promoter with low basal activity
SV40 ori
1918 .. 2053  =  136 bp
SV40 origin of replication
SV40 ori
1918 .. 2053  =  136 bp
SV40 origin of replication
ORF:  2917 .. 3366  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2102 .. 2896  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  111 .. 872  =  762 bp
ORF:  253 amino acids  =  27.6 kDa
ORF:  2274 .. 2660  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  38 .. 580  =  543 bp
ORF:  180 amino acids  =  19.4 kDa
ORF:  599 .. 856  =  258 bp
ORF:  85 amino acids  =  9.4 kDa
ORF:  2411 .. 2947  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3122 .. 3355  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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pLVX-MetLuc Reporter
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pMCS-Cypridina Luc
pSF-CMVe-FLuc
pCMV-Red Firefly Luc
pGL4.39[luc2P ATF6 RE Hygro]
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pCRE-MetLuc2-Reporter
pGL4.40[luc2P MRE Hygro]
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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
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pGL3-Control
pGL4.49[luc2P TCF-LEF RE Hygro]
pMirTarget
pSF-PromMCS-FLuc
pGL3-Enhancer
pGL4.50[luc2 CMV Hygro]
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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
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pSV40-GLuc
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pGL4.72[hRlucCP]
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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]
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pTK-Gaussia-Dura Luc
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pXPG
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