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Luciferase Plasmid Guide


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.

Split-Luciferase and Protein-Protein Interaction Assays

Bioluminescence Resonance Energy Transfer (BRET)

RNA Biology

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 emtpy backbone plasmids used to 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 Creation 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
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 Creation 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
141285 pGWB501NL3F10H NanoLuc® Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants Andrew Millar
141285 pGWB601NL3F10H NanoLuc® Creation of C-terminal luciferase fusions (NanoLuc-3xFLAG-10xHis) using Gateway cloning for expression in plants Andrew Millar
141285 pGWB701NL3F10H 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

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-Akaluc 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
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.

ID Plasmid Description Gene/Insert PI

Do you have suggestions for other plasmids that should be added to this list?

Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!