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 popular tool for scientists investigating the effect of regulatory elements. Elements such as promoters, enhancers and untranslated regions, or RNAi or CRISPR target sites can be inserted into a luciferase-containing plasmid and the effects on gene expression can be measured by the luminescence output.

The most popular types of luciferase found in Addgene’s collection include:

• Firefly luciferase (Fluc)
• Renilla luciferase (Rluc)
• Gaussia luciferase (Gluc)
• NanoLuc® (Nluc)

The mammalian codon-optimized version of Firefly luciferase is often referred to as Luc2. Other luciferases in our collection include the red and green Click-beetle luciferases (CBRluc and CBGluc, respectively), Cypridina luciferase, and Luciola luciferase.

Each type of luciferase has advantages and disadvantages for different applications. Firefly luciferase is considerably brighter than Renilla luciferase, but is more susceptible to enzyme inhibition. Both firefly 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, attributes that have made it popular with scientists since its release.

Note that different luciferase variants require different substrates and detection wavelengths. To learn more about the advantages and uses of different luciferases, check the original publication or read our Luciferase blog post.

Highlighted Luciferase Tools

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.
• teLuc: Red-shifted bioluminescence when paired with diphenylterazine (DTZ).
• 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 and neoLux1.2: Artificial luciferases desined de novo using deep-learning approaches.

Split Luciferase

• NanoBit : The NanoLuc® enzyme is split into Large (LgBiT; 18 kDa) and Small (SmBiT; 11 amino acids) fragments for protein-protein interaction assays. Each protein of interest is fused to either LgBiT or SmBiT. If the two proteins interact, the NanoBiT subunits come together to form an active enzyme and generate luminescence. Find plasmids containing LgBit and SmBit.
• Cluc-FKBP and FRB-Nluc: Split firefly luciferase reporter of rapamycin-inducible interaction.
• nLuc and cLuc: Constructs for split firefly luciferase complementation assays in plants.

Bioluminescence Resonance Energy Transfer (BRET)

• 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.
• NanoBRET: Constructs using NanoLuc® as a BRET energy donor and HaloTag protein labeled with a fluorophore as the acceptor.
• Luciferases with tunable wavelengths: NanoLuc® with either SNAP‐tag or HaloTag in which labeling of the self‐labeling tag with a fluorophore shifts the emission to that of the fluorophore.

RNA Biology

• Splicing reporters from Younis et al., 2010 and Monteys et al., 2021.
• TREAT biosensor that uses Renilla luciferase and a Halo tag to assay mRNA turnover in real time.
• Nonsense-mediated decay reporter from Baird et al., 2018.

Other Sensors and Tools

• Optimized STARR-seq (Self-Transcribing Active Regulatory Region) plasmids: Genome-wide screening of enhancer activity using a firefly luciferase reporter. These plasmids improve on the original STARR-seq by using the bacterial ORI as the core promoter, which was a source of false positives with the original.
• Structurally destabilized firefly luciferases: A set of three increasingly destabilized mutants which serve as sensors of proteome stress.
• pHLuc: An extracellular pH sensor consisting of a pH-sensitive GFP (superecliptic pHluorin), a pH-stable OFP (Antares), and Nanoluc®.
• CalFlux VNT: An intracellular calcium sensor created by fusing NanoLuc® and Venus fluorophore to Troponin C Ca²⁺ binding domain.
• Luminopsins: Luciferase-opsin fusion proteins for bioluminescent chemo-optogenetics.

Find more plasmids including NanoLuc® fusions, constructs for NanoBRET and NanoBiT assays, and more in the Promega Plasmid Collection.

Empty Backbones

Browse or search for popular 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
	18760	MSCV IRES Luciferase	Firefly	Retroviral expression of firefly luciferase and gene of interest	Scott Lowe
	21375	pHIV-Luciferase	Firefly	Lentiviral vector for insertion of your gene of interest upstream of IRES-firefly luciferase.	Bryan Welm
	60323	pGL4.23-GW	Firefly	Insertion of 5' promoter/enhancer regions using Gateway cloning.	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	Dual reporter vector. 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
	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
	64034	pGL4Luc-RLuc	Firefly/Renilla	Dual reporter reporter vector allowing the study of biscistronic promoters.	Daniel Christophe
	119760	pSGDlucV3.0	Firefly/Renilla	Dual reporter vector. Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase. Renilla luciferase is expressed from a T7 promoter for normalization.	John Atkins
	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
	212935	pGL4.84(hRlucCP/Puro)	RapidResponse™ 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 can be used to monitor transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of expression. Browse or search for popular plasmids expressing luciferase. We also offer ready-to-use AAV preparations of select luciferase expression plasmids.

	ID	Plasmid	Luciferase Type	Promoter	Description	PI
	60226	AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR	Renilla	EF1α	AAV-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
	18782	MSCV Luciferase PGK-hygro	Firefly	SV40	Retroviral expression of firefly luciferase	Scott Lowe
	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
	102320	FUW-RLuc-T2A-PuroR	Renilla	Ubc	Lentiviral Expression of Renilla luciferase	Rudolf Jaenisch
	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
	100984	pGL4.18 CMV-Luc	Firefly	CMV	Mammalian expression of firefly luciferase	Lee Helman
	131860	pZmUBQ:LUC	Firefly	ZmUBQ	Plant expression of firefly luciferase from the Zea mays ubiquitin promoter.	Paul Schulze-Lefert
	83282	pAAV-CAG-RLuc	Renilla	CAG	AAV expression of Renilla luciferase	Mark Kay
	83280	pscAAV-CAG-RLuc	Renilla	CAG	Vector for packaging self-complementary AAV expressing 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	Transcriptional unit for firefly luciferase expression in plants (GoldenBraid 2.0).	Diego Orzaez
	68215	pEGB 35S:Renilla:Tnos (GB0109)	Renilla	35S	Transcriptional unit for Renilla luciferase expression in plants (GoldenBraid 2.0).	Diego Orzaez
	118069	MLRV	Multiple	Multiple	Multi-luciferase reporter vector including five transcriptional reporters for NF-kb, TGF-b, c-Myc, p53, and MAPK/JNK plus a constitutive control.	Koen Venken
	133315	pMKV060	Firefly	AtUbi10	BeYDV replicon on T-DNA backbone expressing firefly luc for monitoring plant transfection.	Daniel Voytas
	133312	pMKV057	Firefly	AtUbi10	BeYDV replicon on T-DNA backbone expressing WUS2 and IPT, with firefly luc for monitoring plant transfection.	Daniel Voytas
	181934	pC0037	Gaussia, Cypridina	EF1α, CMV	Dual secreted luciferase reporter (EF1α-Gaussia luciferase, CMV-Cypridina luciferase).	Feng Zhang

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.