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Promoterless NanoLuc® Genetic Reporter Basic Vectors

Insert a Putative Promoter to Drive Expression of NanoLuc® Luciferase

  • Multiple cloning site offers an array of restriction sites for cloning
  • Offers two engineered genes for your reporter assay: Nluc and NlucP
  • NanoLuc® enzyme and its substrate are about 100-fold brighter than other luciferases, producing a high intensity, glow-type luminescence

Choose a luciferase gene

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Catalog number selected: N1001

$ 395.00
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Promoterless NanoLuc® Genetic Reporter Basic Vectors
Nluc/20µg
$ 395.00
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The pNL1.1[Nluc] and pNL1.2[NlucP] Vectors are used to clone a putative promoter to drive expression of the bright NanoLuc® reporter protein.

NanoLuc® (Nluc) luciferase is a small enzyme (19.1kDa) engineered for optimal performance as a luminescent reporter. The enzyme is about 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase using a novel substrate, furimazine, to produce high intensity, glow-type luminescence. The luminescent reaction is ATP-independent and designed to suppress background luminescence for maximal assay sensitivity.

Applications

  • Transcription regulation.
  • Virus-cell interactions.
  • Compound screening.
  • Post-translational modifications.
  • GPCR signaling.
  • Cell signaling.
  • Promoter analysis.

About NanoLuc® Luciferase and the pNL Vectors

For use as a genetic reporter, multiple forms of NanoLuc® luciferase have been configured to meet differing experimental objectives. Unfused Nluc offers maximal light output and sensitivity, and NanoLuc®-PEST (NlucP) closely couples protein expression to changes in transcriptional activity and increased signal-to background ratios. When a secreted reporter is preferred, there is NanoLuc® luciferase fused to an N-terminal secretion signal (secNluc). Luminescence is linearly proportional to the amount of NanoLuc® protein over a 1,000,000-fold concentration range, with a signal half-life ≥2 hours when detected with Nano-Glo® Luciferase Assay Reagent.

NanoLuc® luciferase possesses a number of physical properties that make it an excellent reporter protein:

  • very small, monomeric enzyme (171 amino acids; 513bp)
  • high thermal stability (Tm = 60°C)
  • active over a broad pH range (pH 6–8)
  • no post-translational modifications or disulfide bonds
  • uniform distribution in cells
  • emission spectrum well suited for bioluminescence resonance energy transfer (BRET; λmax = 465nM).

NanoLuc® luciferase is made available in a variety of plasmids designed for use in reporter gene assays of transcriptional control and with each of the NanoLuc® forms (unfused Nluc, PEST destabilized NlucP, and secreted secNluc). The different pNL variations are designed for the following:

  • pNL1: cloning of a known or putative promoter region
  • pNL2: cloning of a known or putative promoter region and establishment of a stable cell line through Hygromycin selection
  • pNL3: cloning of a binding site or response element not in need of a basic promoter (such as are present in the pNL3.2.NF-κB-RE vector)
  • Control plasmids for the unfused, PEST-destabilized and secreted Nluc forms also are available.

The pNL vector series uses a pGL4-based backbone for easy sequence transfer from existing plasmids. This backbone design also reduces anomalous results by removing many transcription factor binding sites and other potential regulatory elements. The Nluc gene variations are codon optimized and have had many potential regulatory elements or other undesirable features removed (such as common restriction enzyme sites).

10321MA-W

pNL1.1[Nluc] Vector GenBank® Accession Number JQ437370 and vector sequence text file.

10322MA-W

pNL1.2[NlucP] Vector GenBank® Accession Number JQ437371 and vector sequence text file.

A comparison of light output and response dynamics between NanoLuc® and firefly luciferase constructs.

A comparison of light output and response dynamics between NanoLuc® and firefly luciferase constructs.
A comparison of light output and response dynamics between NanoLuc® and firefly luciferase constructs.

NanoLuc® Technology performs similarly to firefly luciferase but gives higher light output.

NanoLuc® Technology performs similarly to firefly luciferase but gives higher light output.
NanoLuc® Technology performs similarly to firefly luciferase but gives higher light output.

Specifications

You are viewing: N1001 Change Configuration

What's in the box?

Item Part # Size
pNL1.1[Nluc] Vector N100A 1 × 20μg

SDS

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Download SDSPDF (41 KB) – English (United States)

Certificate of Analysis

See all certificates for N1001

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researchers may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product or derivatives by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researcher may (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence; and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use of this product or derivatives is authorized without the prior express written consent of Promega.

In addition, researchers must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product and its derivatives; or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.

For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(2a) use NanoBRET™-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives; or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.

Researchers may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730 and 9,951,373 and other patents and patents pending.

Specifications

You are viewing: N1011 Change Configuration

What's in the box?

Item Part # Size
pNL1.2[NlucP] Vector N101A 1 × 20μg

SDS

Choose language:

Download SDSPDF (41 KB) – English (United States)

Certificate of Analysis

See all certificates for N1011

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researchers may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product or derivatives by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researcher may (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence; and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use of this product or derivatives is authorized without the prior express written consent of Promega.

In addition, researchers must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product and its derivatives; or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.

For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(2a) use NanoBRET™-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives; or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.

Researchers may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730 and 9,951,373 and other patents and patents pending.

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