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Control NanoLuc® Reporter Vector with Hygromycin Selection

Vector that Expresses NanoLuc® Luciferase Using the CMV Promoter

  • Used as a control vector for the NanoLuc® fusion vectors
  • Includes an optimized NanoLuc® luciferase gene that codes for a reporter protein with a shorter half-life (NlucP)
  • Hygromycin resistance selectable marker means choosing transient transfection or generating a stable cell line

Size

Catalog number selected: N1411

$ 395.00
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Control NanoLuc® Reporter Vector with Hygromycin Selection
20μg
$ 395.00
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The pNL3.2.CMV Vector constitutively expresses the NanoLuc® reporter fused to a PEST destabilization domain (NlucP). The NlucP fusion protein naturally accumulates at low intracellular levels due to constitutive proteosomal degradation, thus serving as a negative control for experiments configured to measure regulated changes in NanoLuc® luciferase expression levels (e.g., NanoLuc® fusion vectors). This vector also encodes a hygromycin resistance gene for selection in mammalian cells.

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.

About the NanoLuc® Luciferase Reporter Enzyme

NanoLuc® luciferase in the form of NanoLuc®-PEST (NlucP) closely couples protein expression to changes in transcriptional activity and increased signal-to background ratios. The PEST sequence appended to Nluc destabilizes the reporter protein, make NanoLuc® luciferase more responsive to changes in the cell. 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).

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 NlucP gene is codon optimized and has had many potential regulatory elements or other undesirable features removed (such as common restriction enzyme sites).

Sequence Information

pNL3.2.CMV Vector GenBank® Accession Number KF853603 and vector sequence text file.

Protocols

Specifications

What's in the box?

Item Part # SizeConcentration

pNL3.2.CMV Vector

N141A 1 × 20μg1μg/μl

SDS

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

Certificate of Analysis

See all certificates for N1411

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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