pNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]

Vector with a minimal promoter and NF-κB response element for studying cell signaling using destabilized NanoLuc® luciferase.

Sequence Author: Promega

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HomePlasmidsLuciferase VectorspNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]
AarI - BfuAI - BspMI (4882) RVprimer3 (4861 .. 4880) poly(A) signal BsmBI (4709) SpeI (4696) BsaHI (4404) BstZ17I (4377) SacII (4261) PvuI (4237) BstEII (3792) BstXI - PstI (3789) AleI (3787) NotI (3765) BspHI (3665) AlwNI (3361) ApaLI (3259) BciVI (3148) PciI (2945) RVprimer4 (2746 .. 2765) SalI (2695) BstBI (2681) AgeI (2616) PmeI (2605) BglI - SfiI (8) Acc65I (14) KpnI (18) Eco53kI (23) SacI (25) NheI (27) BmtI (31) BglII (85) BglI - SfiI (98) HindIII (104) PflFI - Tth111I (243) EcoNI (252) PpuMI - SanDI (571) EcoRI (692) FseI (847) PsiI (968) HpaI (988) MfeI (997) BamHI (1090) PvuII (1200) SexAI * (1291) BseRI (1520) StuI (1523) AvrII (1524) BsaAI (1755) PspOMI (1978) ApaI (1982) BssHII (2580) pNL3.2.NF- κ B-RE[NlucP/NF- κ B-RE/Hygro] 4912 bp
AarI  (4882)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
BfuAI  (4882)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (4882)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BsmBI  (4709)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
SpeI  (4696)
1 site		 A C T A G T T G A T C A
BsaHI  (4404)
1 site		 G R C G Y C C Y G C R G
BsaHI is typically used at 37°C, but is even more active at 60°C.
BstZ17I  (4377)
1 site		 G T A T A C C A T A T G
SacII  (4261)
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.
PvuI  (4237)
1 site		 C G A T C G G C T A G C
BstEII  (3792)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BstXI  (3789)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
PstI  (3789)
1 site		 C T G C A G G A C G T C
AleI  (3787)
1 site		 C A C N N N N G T G G T G N N N N C A C
NotI  (3765)
1 site		 G C G G C C G C C G C C G G C G
BspHI  (3665)
1 site		 T C A T G A A G T A C T
AlwNI  (3361)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
ApaLI  (3259)
1 site		 G T G C A C C A C G T G
BciVI  (3148)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
PciI  (2945)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
SalI  (2695)
1 site		 G T C G A C C A G C T G
BstBI  (2681)
1 site		 T T C G A A A A G C T T
AgeI  (2616)
1 site		 A C C G G T T G G C C A
PmeI  (2605)
1 site		 G T T T A A A C C A A A T T T G
BglI  (8)
2 sites		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
SfiI  (8)
2 sites		 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.
Acc65I  (14)
1 site		 G G T A C C C C A T G G
KpnI  (18)
1 site		 G G T A C C C C A T G G
Eco53kI  (23)
1 site		 G A G C T C C T C G A G
SacI  (25)
1 site		 G A G C T C C T C G A G
NheI  (27)
1 site		 G C T A G C C G A T C G
BmtI  (31)
1 site		 G C T A G C C G A T C G
BglII  (85)
1 site		 A G A T C T T C T A G A
BglI  (98)
2 sites		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
SfiI  (98)
2 sites		 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.
HindIII  (104)
1 site		 A A G C T T T T C G A A
PflFI  (243)
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  (243)
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.
EcoNI  (252)
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.
PpuMI  (571)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
SanDI  (571)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different SanDI sites may not be compatible.
EcoRI  (692)
1 site		 G A A T T C C T T A A G
FseI  (847)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
PsiI  (968)
1 site		 T T A T A A A A T A T T
HpaI  (988)
1 site		 G T T A A C C A A T T G
MfeI  (997)
1 site		 C A A T T G G T T A A C
BamHI  (1090)
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.
PvuII  (1200)
1 site		 C A G C T G G T C G A C
SexAI  (1291)
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.
BseRI  (1520)
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  (1523)
1 site		 A G G C C T T C C G G A
AvrII  (1524)
1 site		 C C T A G G G G A T C C
BsaAI  (1755)
1 site		 Y A C G T R R T G C A Y
PspOMI  (1978)
1 site		 G G G C C C C C C G G G
ApaI  (1982)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BssHII  (2580)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
RVprimer3
20-mer  /  50% GC
1 binding site
4861 .. 4880  =  20 annealed bases
Tm  =  54°C
RVprimer4
20-mer  /  65% GC
1 binding site
2746 .. 2765  =  20 annealed bases
Tm  =  62°C
HygR
1570 .. 2607  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
1570 .. 2607  =  1038 bp
345 amino acids  =  38.4 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
AmpR
3794 .. 4654  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3794 .. 4585  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3794 .. 4654  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4586 .. 4654  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3794 .. 4654  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
Nluc
180 .. 692  =  513 bp
171 amino acids  =  19.1 kDa
Product: NanoLuc® luciferase
human codon-optimized
Nluc
180 .. 692  =  513 bp
171 amino acids  =  19.1 kDa
Product: NanoLuc® luciferase
human codon-optimized
hPEST
696 .. 815  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
hPEST
696 .. 815  =  120 bp
40 amino acids  =  4.2 kDa
Product: PEST degradation sequence from mouse ornithine decarboxylase
human codon-optimized
ori
3006 .. 3594  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3006 .. 3594  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
SV40 promoter
1182 .. 1539  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1182 .. 1539  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
867 .. 988  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
867 .. 988  =  122 bp
SV40 polyadenylation signal
pause site
4821 .. 4912  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
pause site
4821 .. 4912  =  92 bp
RNA polymerase II transcriptional pause signal from the human α2 globin gene
NF-κB RE
33 .. 84  =  52 bp
NF-κB response element (5 copies)
NF-κB RE
33 .. 84  =  52 bp
NF-κB response element (5 copies)
poly(A) signal
2631 .. 2679  =  49 bp
synthetic polyadenylation signal
poly(A) signal
2631 .. 2679  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4759 .. 4807  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4759 .. 4807  =  49 bp
synthetic polyadenylation signal
minP
116 .. 147  =  32 bp
minimal TATA-box promoter with low basal activity
minP
116 .. 147  =  32 bp
minimal TATA-box promoter with low basal activity
SV40 ori
1390 .. 1525  =  136 bp
SV40 origin of replication
SV40 ori
1390 .. 1525  =  136 bp
SV40 origin of replication
ORF:  1570 .. 2607  =  1038 bp
ORF:  345 amino acids  =  38.4 kDa
ORF:  129 .. 818  =  690 bp
ORF:  229 amino acids  =  25.1 kDa
ORF:  3924 .. 4190  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4836 .. 181  =  258 bp
ORF:  85 amino acids  =  9.2 kDa
ORF:  3794 .. 4654  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1903 .. 2328  =  426 bp
ORF:  141 amino acids  =  14.3 kDa
ORF:  108 .. 443  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
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Individual Sequences & Maps

pBI-CMV5
pGL4.22[luc2CP Puro]
pGL4.81[hRlucCP Neo]
pRL-null
pBIND-ER(alpha)
pGL4.23[luc2 minP]
pGL4.82[hRluc Puro]
pRL-SV40
pBIND-GR
pGL4.24[luc2P minP]
pGL4.83[hRlucP Puro]
pRL-TK
pCBG68-Basic
pGL4.25[luc2CP minP]
pGL4.84[hRlucCP Puro]
pSELECT-zeo-Lucia
pCBG68-Control
pGL4.26[luc2 minP Hygro]
pGLuc Mini-TK 2
pSF-CMV-EMCV-FLuc
pCBG99-Basic
pGL4.27[luc2P minP Hygro]
pGLuc-Basic 2
pSF-CMV-EMCV-RLuc
pCBG99-Control
pGL4.28[luc2CP minP Hygro]
phRG-B
pSF-CMV-FLuc
pCBR-Basic
pGL4.29[luc2P CRE Hygro]
phRG-TK
pSF-CMV-FMDV-FLuc
pCBR-Control
pGL4.30[luc2P NFAT-RE Hygro]
phRL-CMV
pSF-CMV-FMDV-RLuc
pCLuc Mini-TK 2
pGL4.31[luc2P GAL4UAS Hygro]
phRL-null
pSF-CMV-PGK-FLuc
pCLuc-Basic 2
pGL4.32[luc2P NF-kB-RE Hygro]
phRL-SV40
pSF-CMV-PGK-RLuc
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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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