Luciferase Plasmid Collection

Luciferase is the enzyme responsible for the bioluminescence found in a diverse number of organisms, ranging from bacteria to aquatic animals to insects. Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust signal, sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors.

The most common types of luciferase found in Addgene’s collection are Firefly luciferase (Fluc), Renilla luciferase (Rluc), and NanoLuc® Luciferase (Nluc). Luc2 is the second-generation version of Firefly luciferase that has been codon optimized for expression in mammalian systems. Other luciferases in our collection include Gaussia luciferase (Gluc) and the red and green Click-beetle luciferases (CBRluc and CBGluc, respectively). Like Firefly luciferase, Click-beetle red luciferase has also been codon optimized (CBR2opt) for mammalian expression systems.

Each type of luciferase has advantages and disadvantages depending on the application. Firefly luciferase is considerably brighter than Renilla luciferase, but is more susceptible to enzyme inhibition. Both Firefly luciferase and Renilla luciferase have short protein half-lives, making them useful as transcriptional reporters. However, they are not secreted, so cells must be lysed or the substrate must be delivered intracellularly in order to measure luminescence. Gaussia luciferase is secreted and more stable, making time course experiments possible, but its short luminescence half-life makes it too dim for many experimental systems. Firefly and Renilla luciferase are less-than-ideal protein tags due to their large size. NanoLuc® Luciferase is small, bright, and a secreted version of the protein is available – these attributes have made it popular with scientists since its release.

Luciferase-containing plasmids are commonly used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these regulatory elements on gene expression. Empty backbones for inserting regulatory elements before or after the luciferase ORF can be found in a table below, as well as plasmids that already contain a region of interest before or after luciferase. In addition, Addgene has many luciferase expression constructs available that are driven by a strong constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. Scientists have developed a wide range of luciferase-based plasmid systems for many different applications. Browse the Collection Highlights section to learn more.

Highlights of Luciferase Tools in our Collection

Modified/Novel Luciferases

• AkaLuc: A modified firefly luciferase engineered to use the red-shifted substrate Akalumine with significantly brighter signal and emission in the near-infrared range, enabling enhanced bioluminescence imaging in vivo.
• Luciferases with tunable wavelengths: NanoLuc® with either SNAP‐tag or HaloTag7 tag in which labeling of the self‐labeling tag with a fluorophore shifts the emission maximum of NanoLuc to that of the fluorophore.
• Conformationally destabilized Firefly luciferase mutants: A set of three sensors that are increasingly structurally destabilized versions of Firefly luciferase which serves as a convenient toolkit to assess the proteostasis status in a wide range of experimental systems.
• LumiLuc and LumiScarlet: LumiLuc and LumiScarlet are ATP-independent luciferases derived from teLuc that emit yellow/green light and red light, respectively, for increased usefulness in in vivo applications.
• LuxSit-i: An artificial luciferase desined de novo using deep-learning approaches.

Split-Luciferase and Protein-Protein Interaction Assays

• Cluc-FKBP, FRB-Nluc: Split Firefly luciferase to assess FKBP and FRP interactions in the mTOR signaling pathway in vivo.
• DULIP (DUal Luminescence-based Co-Immunoprecipitation) plasmids: Luminescence-based protein-protein screening method for the systematic and quantitative analysis of co-immunoprecipitations using both Renilla and Firefly luciferase. Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016 blog post.
• LuTHy plasmids (Double‐Readout Luminescence‐based Two‐Hybrid): Combines in vivo BRET with ex vivo luminescence-based co-immunoprecipitation to assess protein-protein interactions.
• NanoLuc® complementation: Protein-protein assays based on a split NanoLuc® have been used to assess GPCR-G protein interactions, protein dimerization, endosomal escape of cell-penetrating peptides, and more.
• nLuc/cLuc for split Firefly Luciferase complementation in plants.

Bioluminescence Resonance Energy Transfer (BRET)

• CalFlux VNT: An intracellular calcium sensor created by fusing NanoLuc® and Venus fluorophore to Troponin C Ca²⁺ binding domain.
• Nano-lanterns, Enhanced Nano-lanterns, and Antares: Fluorescent proteins are fused to Renilla luciferase (Nano-lanterns) or NanoLuc® (Antares and Enhanced Nano-lanterns) to allow for BRET-based in vivo imaging of multiple cellular processes, biosensors, or optogenetic tools.
• pcDNA3.1+-pHlash, pRSETb-pHlash: A pH sensor created by fusing a bright mutant of Renilla luciferase (Rluc8) through an Ala-Glu-Leu linker to the circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot Plasmid Technologies from 2015 blog post.

RNA Biology

• CMV-LUC2CP/intron/ARE, CMV-LUC2CP/ARE: A splicing reporter that contains an intron in the middle of the Firefly luciferase gene..
• TREAT (3(Three)′-RNA End Accumulation during Turnover): Single-molecule mRNA turnover biosensor that utilizes Renilla luciferase and a Halo tag to assay mRNA decay in real time.
• pKC-4.04, pKC-4.06: Nonsense-mediated decay reporter and control plasmid utilizing Firefly luciferase.

Transcriptional Element Screening

• Optimized STARR-seq (Self-Transcribing Active Regulatory Region) plasmids: Genome-wide screening of enhancer activity using a Firefly luciferase reporter. These plasmids improve over the original STARR-seq plasmids by using the bacterial ORI as the core promoter, which was a source of false positives with the original STARR-seq method. Learn more about STARR-seq in our 27 Hot Plasmids from 2016 blog post.
• pGL3-lRF, pGL3-LRR: Genomic tiles spanning a region of interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of transcriptional regulatory region in specific cell types. Used to identify functional SNPs from loci determined to be important in disease.

Empty Backbones

This table is a collection of empty backbone plasmids into which you can clone your regulatory element or gene of interest into to create a luciferase reporter. Unless otherwise noted, these plasmids are meant for expression in mammalian systems.

	ID	Plasmid	Luciferase Type(s)	Description	PI
	16539	pBV-Luc	Firefly	Insertion of 5' promoter/enhancer regions	Bert Vogelstein
	12178	pIS0	Firefly	Insertion of target sequence in the 3' UTR	David Bartel
	12179	pIS1	Renilla	Insertion of target sequence in the 3' UTR	David Bartel
	12177	pIS2	Renilla	Insertion of target sequence in the 3' UTR	David Bartel
	101139	pCMV-IRES-Renilla Luciferase-IRES-Gateway-Firefly Luciferase (pIRIGF)	Firefly/Renilla	Creating ORF-Firefly luciferase gene fusions. Renilla luciferase under the control of a CMV promoter is present for normalization	William Kaelin
	71248	pXPG	Firefly	Insertion of 5' promoter/enhancer regions. Mutations within the luciferase gene allow for increased copy number in E. coli and expression of luciferase in the cytoplasm of mammalian cells	Peter Cockerill
	64784	pGL3-Basic-IRES	Firefly	Insertion of 5' promoter/enhancer regions	Joshua Mendell
	212936	pGL3 Basic Vector	Firefly	Vector for investigating regions controlling transcription	Debrya Groskreutz
	60323	pGL4.23-GW	Firefly	Insertion of 5' promoter/enhancer regions using Gateway cloning. Lentiviral.	Jorge Ferrer
	87067	pcDNA3.1-ccdB-Nanoluc	NanoLuc®	Creation of N-terminal Nanoluc fusions using Gateway cloning	Mikko Taipale
	87075	pLenti6.2-ccdB-Nanoluc	NanoLuc®	Creation of C-terminal Nanoluc fusions using Gateway cloning. Lentival.	Mikko Taipale
	87078	pLenti6.2-Nanoluc-ccdB	NanoLuc®	Creation of N-terminal Nanoluc fusions using Gateway cloning. Lentiviral.	Mikko Taipale
	12563	PCD FL0X	Firefly	Insertion of imperfect miRNA sequence in the 3' UTR	Carl Novina
	33154	pAC-Luc	Firefly	Creation of N-terminal Firefly fusions in Drosopholia	Michael Rosbash
	128046	pGL3basic luciferase and renilla_polyA	Firefly/Renilla	Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase. Renilla luciferase is co-expressed.	Oskar Laur
	87070	pcDNA3.1-Nanoluc-ccdB	NanoLuc®	Creation of N-terminal Nanoluc fusions using Gateway cloning	Mikko Taipale
	87065	pcDNA3.1-ccdB-Renilla	Renilla	Creation of C-terminal Renilla luciferase fusions using Gateway cloning	Mikko Taipale
	87066	pcDNA3.1-ccdB-Firefly	Firefly	Creation of C-terminal Firefly luciferase fusions using Gateway cloning	Mikko Taipale
	87068	pcDNA3.1-Renilla-ccdB	Renilla	Creation of N-terminal Renilla luciferase fusions using Gateway cloning	Mikko Taipale
	87069	pcDNA3.1-Firefly-ccdB	Firefly	Creation of N-terminal Firefly luciferase fusions using Gateway cloning	Mikko Taipale
	87074	pLenti6.2-ccdB-Firefly	Firefly	Creation of C-terminal Firefly luciferase fusions using Gateway cloning. Lentiviral.	Mikko Taipale
	87077	pLenti6.2-Firefly-ccdB	Firefly	Creation of N-terminal Firefly luciferase fusions using Gateway cloning. Lentiviral.	Mikko Taipale
	87789	pPTluc	Firefly	Insertion of 5' promoter/enchancer regions in Drosopholia	Herman Wijnen
	126033	pTETRIS-cargo	Firefly	For use as a reporter of the ability of a given lncRNA to cis-repress	Mauro Calabrese
	106253	pLS-mP-Luc	Firefly	Insertion of 5' promoter/enhancer regions. Lentiviral	Nadav Ahituv
	106292	pLS-SV40-mP-Rluc	Renilla	Insertion of 5' promoter/enhancer regions. Lentiviral	Nadav Ahituv
	108228	pGreen2x35SLUC	Firefly/Renilla	Creating ORF-Firefly luciferase gene fusions in plants. Renilla luciferase under the control of a CMV promoter is present for normalization	Alejandro Ferrando
	114670	pGL3-TK-5UTR-BsmBI-Luciferase	Firefly	Insertion of 5' promoter/enhancer regions. Blue/white screening can be used for screening of transformants in cloning process.	Lynne Postovit
	119760	pSGDlucV3.0	Firefly/Renilla	Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase. Renilla luciferase is expressed from a T7 promoter for normalization. FMDV StopGo sequences flank the MCS for improved normalization.	John Atkins
	167576	mpCHECK2	Firefly/Renilla	Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly luciferase expression under the control of a HSV TK promoter for normalization	Ting Ni
	174050	pGWB-nLUC	Firefly	Creation of N-terminal Firefly luciferase fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174051	Alberto Macho
	174051	pGWB-cLUC	Firefly	Creation of C-terminal Fifrefly luciferase fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174050	Alberto Macho
	136010	psi-CHECK3	Firefly/Renilla	Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly luciferase expression under the control of a HSV TK promoter for normalization. Modified from psi-CHECK2 so both Renilla and Firefly luciferase contain introns	Anthony Leung
	141285	pGWB401NL3F10H	NanoLuc®	Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants	Andrew Millar
	159707	pG20_LUC2_Hyg	Firefly	Creation of C-terminal Firefly luciferase fusion for expression in plants	Hans-Henning Kunz
	159708	pG20_LUC2_Blp	Firefly	Creation of C-terminal Firefly luciferase fusion for expression in plants	Hans-Henning Kunz
	141305	pRATIO2251	Firefly	Creation of C-terminal Firefly luciferase fusion for expression in plants, co-expressed with mScarlet-I for ratiometric imaging	David Nelson
	141301	pRATIO1267	RedLuc/Gaussia	Creation of C-terminal Red Luciferase fusion for expression in plants, co-expressed with Gaussia luciferase for ratiometric imaging	David Nelson
	212935	pGL4.84(hRlucCP/Puro)	RapidResponse(TM) Renilla	Vector for investigating regions controlling transcription	Pete Stecha
	212933	pGL4.82(hRluc/Puro)	Renilla	Vector for investigating regions controlling transcription	Pete Stecha

Expression Constructs

Luciferase driven by a strong constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. Use this table to find plasmids expressing luciferase. We also offer ready-to-use AAV preparations of select luciferase expression plasmids; these are noted with the viral service icon .

	ID	Plasmid	Luciferase Type	Promoter	Description	PI
	60226	AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR	Renilla	EF1α	AAV9-CRISPR knockout plasmid. Renilla luciferase is used to monitor viral transfection	Feng Zhang
	21471	pLenti PGK V5-LUC Neo (w623-2)	Firefly	PGK	Lentiviral expression of firefly luciferase	Eric Campeau
	21474	pLenti CMV V5-LUC Blast (w567-1)	Firefly	CMV	Lentiviral expression of firefly luciferase	Eric Campeau
	18964	Luciferase-pcDNA3	Firefly	CMV	Mammalian expression of firefly luciferase	William Kaelin
	21375	pHIV-Luciferase	Firefly	EF1α/IRES	Lentiviral expression of firefly luciferase and gene of interest	Bryan Welm
	18760	MSCV IRES Luciferase	Firefly	(IRES)	Retroviral expression of firefly luciferase and gene of interest	Scott Lowe
	18782	MSCV Luciferase PGK-hygro	Firefly	SV40	Retroviral expression of firefly luciferase	Scott Lowe
	14080	pAKlux2	Lux Operon		Bacterial expression of luciferase from broad host range plasmid	Attila Karsi
	105621	Lenti-luciferase-P2A-Neo	Firefly	EF1α	Lentiviral expression of firefly luciferase	Christopher Vakoc
	33307	Ubc.Luc.IRES.Puro	Firefly	Ubc	Lentiviral expression of firefly luciferase	Linzhao Cheng
	74444	pLenti.PGK.blast-Renilla_Luciferase	Renilla	PGK	Lentiviral expression of renilla luciferase	Reuben Shaw
	48688	pUltra-Chili-Luc	Firefly	Ubc	Lentiviral expression of firefly luciferase and dTomato. A gene of interest can also be inserted into the plasmid.	Malcolm Moore
	51819	Syn-ATP	Firefly (modified)	CMV	Optical reporter of presynaptic ATP	Timothy Ryan
	24348	pAC-hRluc	Renilla	Actin5C	Insect expression of renilla luciferase	Liqun Luo
	51970	Nano-lantern/pcDNA3	Nano-lantern	CMV	Mammalian expression of Nano-lantern	Takeharu Nagai
	87121	pcDNA-RLuc8	Renilla	CMV	Mammalian expression renilla luciferase mutant Rluc8, which has increased stability and light output	Sanjiv Sam Gambhir
	104587	pHIV-iRFP720-E2A-Luc	Firefly	EF1α	Lentiviral expression of firefly luciferase and iRFP720 from a bicistronic promoter	Antonius Plagge
	118017	pLX313-Firefly luciferase	Firefly	EF1α	Lentiviral expression of firefly luciferase	William Hahn, David Root
	118016	pLX313-Renilla luciferase	Renilla	EF1α	Lentiviral expression of renilla luciferase	William Hahn, David Root
	105533	pAAV.CMV.Luc.IRES.EGFP.SV40	Firefly	CMV	AAV expression of firefly luciferase and GFP	James Wilson
	22522	phGluc	Gaussia	EF1α	Expression of Gaussia luciferase; GFP is expressed if cells are infected with virus	Christopher Buck
	98580	pLX304 Luciferase-V5 blast	Firefly	CMV	Lentiviral expression of firefly luciferase with a V5 tag	Kevin Janes
	83281	pAAV-CAG-FLuc	Firefly	CAG	AAV expression of firefly luciferase	Mark Kay
	105532	pAAV.CMV.ffLuciferase.SV40	Firefly	CMV	AAV expression of firefly luciferase	James Wilson
	101156	T7-CMVtrans-FFLuc-polyA	Firefly	T7	Expression of firefly luciferase from a cell-free mammalian cell lysate	Marcel Bruchez
	108542	pLenti-EF1a-Luciferase-IRES-Blast-WPRE	Firefly	EF1α	Lentiviral expression of firefly luciferase	Javier Alcudia
	113442	pcDNA3.1 NL	NanoLuc®	CMV	Mammalian expression of Nanoluc with a N-terminal Myc tag	Erich Wanker
	124701	pLenti-PGK-Venus-Akaluc (neo)	Akaluc	hPGK	Lentiviral expression of Venus-Aka-luciferase	Roland Friedel
	120798	pRRL Luciferase	Firefly	hPGK	Lentiviral expression of firefly luciferase	Paul Khavari
	109395	EFS-Vector	Firefly	EF1α	AAV-CRISPR knockout plasmid. Firefly luciferase is used to moniter viral transfection	Sidi Chen
	109396	TBG-Vector	Firefly	TBG	AAV-CRISPR knockout plasmid. Firefly luciferase is used to moniter viral transfection	Sidi Chen
	118412	ssAAV-EF1a-FLuc-WPRE-HgHpA_bac_293	Firefly	EF1α	AAV expression of firefly luciferase	Nicole Paulk
	98294	pF CAG luc IRES neo	Firefly	CAG	Lentiviral expression of firefly luciferase	Brent Stringer
	102320	FUW-RLuc-T2A-PuroR	Renilla	Ubc	Lentiviral Expression of renilla luciferase	Rudolf Jaenisch
	105538	pENN.AAV.TBG.PI.ffLuciferase.RBG	Firefly	TGB	AAV expression of firefly luciferase and GFP	James Wilson
	106457	pCR3.1-Luc	Firefly	CMV	Mammalian expression of firefly luciferase	Georg Weber
	113450	pLenti NL	NanoLuc®	Ubc	Lentiviral expression of Nanoluc with a N-terminal Myc tag	Erich Wanker
	115352	pFL-SV40	Firefly	SV40	Mammalian expression of firefly luciferase	Ming-Chih Lai
	115366	p CIneo-RL	Renilla	CMV	Mammalian expression of humanized renilla luciferase	Witold Filipowicz
	100984	pGL4.18 CMV-Luc	Firefly	CMV	Mammalian expression of firefly luciferase	Lee Helman
	123655	pKM19	Firefly	pIEx	Insect expression of firefly luciferase	Kevin Maringer
	131860	pZmUBQ:LUC	Firefly	CaMV	Zea mays expression of firefly luciferase	Paul Schulze-Lefert
	83282	pAAV-CAG-RLuc	Renilla	CAG	AAV expression of renilla luciferase	Mark Kay
	140328	pLenti-PGK-Venus-Fluc (puro)	Firefly	hPGK	Lentiviral expression of Venus-firefly luciferase	Roland Friedel
	170575	pCMV-FLuc	Firefly	CMV	Retroviral expression of firefly luciferase	David Nemazee
	68216	pEGB 35S:Luciferase:Tnos (GB0110)	Firefly	35S	Expression of firefly luciferase in plants	Diego Orzaez
	68215	pEGB 35S:Renilla:Tnos (GB0109)	Renilla	35S	Expression of Renilla luciferase in plants	Diego Orzaez
	118069	MLRV	Multiple	Multiple	Multi-luciferase reporter vector including transcriptional reporters for NF-kb, TGF-b, c-Myc, p53 and MAPK/JNK transcriptional reporters	Koen Venken
	127223	pMOD_A5801	Firefly	35S	Expression of firefly luciferase in plants	Daniel Voytas
	127224	pMOD_A5803	Firefly	CmYLCV	Expression of firefly luciferase in plants	Daniel Voytas

Reporter Constructs

Already know what gene or regulatory element that you need a luciferase reporter for? Browse or use the search box to find a construct that contains your regulatory element of interest.