 | | | | pNEBR-X1GLuc Control Plasmid | | | |
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Prices are in US dollars and valid only for US orders.
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- Expresses a secreted luciferase allowing for activity assay by sampling cell culture media.
- GLuc cDNA is cloned in the HindIII/NotI sites of pNEBR-X1 Vector.
- GLuc expression is under control of a minimal promoter and 5 copies of GAL4 RE.
- Can be used as a positive control for any expression system that employs a GAL4 DNA Binding Domain activator protein.
- When used in conjunction with pNEBR-R1 Regulator Plasmid expression of GLuc is controlled by the addition of RSL1 synthetic ligand.
- Ampicillin resistance for propagation in E. coli.
Description:
pNEBR-X1GLuc Control Plasmid contains the cDNA encoding Gaussia princeps secreted luciferase (GLuc) cloned into pNEBR-X1 (NEB #N8078), placing it under control of 5 copies of the GAL4 RE and a minimal promoter. The codon usage has been optimized for expression in mammalian cells.
The plasmid can be used as a positive control in the RheoSwitch® Mammalian Inducible Expression System (NEB #E3000). Upon co-transfection into mammalian cells with pNEBR-R1 Regulator Plasmid (NEB #N8079), expression of GLuc is induced by the addition of the RSL1 synthetic ligand (NEB #E3301).
GLuc is a secreted, stable protein allowing for the assay of cell culture media.
Source:
pNEBR-X1Gluc is isolated from E. coli strain ER2272 by a standard plasmid purification procedure.
Concentration:
500 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
pNEBR-X1GLuc Control Plasmid map. Gaussia Luciferase gene (red) is inserted in the HindIII and NotI sites of pNEBR-X1 Vector, placing it under control of the GAL4 RE and minimal promoter (green).
References
- RheoSwitch Mammalian Inducible Expression System Manual., unpublished observations.
- Palli, S.R., Kapitskaya, M.Z., Kumar, M.B. and Cress, D.E. (2003) Eur. J. Biochem., 270, 1308-1315.
- Dhadialla, T.S., Carlson, G.R. and Le, D.P. (1998) Annu. Rev. Entomol., 43, 545-569.
- Kumar, M.B., Potter, D.W., Hormann, R.E., Edwards, A., Tice, C.M., Smith, H.C., Dipietro, M.A., Polley, M., Lawless, M., Wolohan, P.R., Kethidi, D.R. and Palli, S.R. (2004) J. Biol. Chem., 279, 27211-27218.
- Tannous, B.A., Kim, D.E., Fernandez, J.L., Weissleder, R. and Breakefield, X.O. (2005) Mol. Ther., 11, 435-443.
Companion Products
Gaussia Luciferase Assay Kit
Luciferase Cell Lysis Buffer
pCMV-GLuc Control Plasmid
pGLuc-Basic Vector
pGLuc-Reverse Sequencing Primer (24-mer)
pNEBR-R1 Regulator Plasmid
pNEBR-X1 Vector
pNEBR-X1Hygro Vector
RheoSwitch® Ligand RSL1
RheoSwitch® Mammalian Inducible Expression System
TransPass™ D1 Transfection Reagent
TransPass™ D2 Transfection Reagent
Legal
Licenses/Patents/Disclaimers:
By opening this package or by using the materials enclosed within, recipient is legally bound and accepts the following terms and conditions:
Non-Commercial entities: This product is covered by US Patents No. 6,232,107 and other patents that are the legal property as assigned to Prolume Ltd./Nanolight Technologies. This product is licensed only to the purchasing laboratory-research group. Recipient agrees not to transfer this plasmid or derivatives of this vector to any other laboratory, person or research group, even if within the same institution.
Recipient agrees not to alter or make any changes to the nucleotide coding sequence or secretory coding sequence of the luciferase(s) contained within without prior written permission from Prolume Ltd./Nanolight Technologies (www.nanolight.com). Recipient agrees not to file for any patent rights or to any inventions claiming any portion of the luciferase(s) within this material without prior written permission from Prolume Ltd./Nanolight Technologies.
Commercial For-Profit Entities & Non-Profit Foundations (herein referred to as Commercial Recipients): Commercial Recipients wishing to derive products, engage in the sale or license of any products, discover drugs, or make inventions by use of the material enclosed, fully agree to the terms mentioned above for Non-Commercial entities: AND ADDITIONALLY agree to and are hereby bound to use the materials FOR EVALUATION PURPOSES ONLY. Commercial Recipient hereby agrees to destroy and cease use of any materials or derivatives containing any portion of these materials within 180 days from receipt. Commercial Recipient agrees not to use the materials for any use, other than the 180 day suitability evaluation without prior written permission or obtaining a valid license from Prolume Ltd./Nanolight Technologies.
Any recipient that does not accept the license terms mentioned above, shall return the unopened package and materials to NEB for a full refund.
This product is the subject of one or more of U.S. Patent Nos. 5,514,578, 6,245,531, including the corresponding foreign patents (Australian Patent No. 752699, Canadian Patent App No. 2076386, European Patent No. 0517805) and patent applications, U.S. Patent Nos. 6,258,603, 5,530,028 and U.S. Patent Applications 09/965,703, 10/239,134, 10/468,199, 10/468,193, licensed to New England Biolabs.
The purchase of this product conveys to the non-commercial purchaser the non-transferable right to use the purchased amount of the product and components of the product for research use only conducted by the purchaser. For commercial purchasers using this product for research, the purchase of this product conveys to the purchaser the non-transferable right to use the purchased amount of the product and components of the product for a six(6) month evaluation period after the date of purchase for research use only. After the six(6) month evaluation period, the purchaser must contact RheoGene, Inc. to obtain a commercial research license. The purchaser cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for commercial purposes. The purchaser cannot use this product or its components in whole plants, plant cells or humans. The purchaser cannot directly or indirectly disassemble, decompile, translate, reverse engineer, modify or adapt, (a) this product or (b) its components. The purchaser may transfer information or materials made through the use of this product to a scientific collaborator, provided that such transfer is not for the commercial purposes of the purchaser, and that such collaborator agrees in writing (a) not to transfer such materials to any third party, and (b) to use such transferred materials and/or information solely for research and not for commercial purposes.
Commercial purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data (3) use of the products or its components for therapeutic, diagnostic or prophylactic purposes; (4) use of the product or its components to screen for agonists to the RheoSwitch® System; or (5) resale of the product or its components, whether or not such product or its components are resold for use in research. If the purchaser is not willing to accept the limitations of the limited use statement, NEB is willing to accept return of the products with a full refund. For information on commercial use of this product or obtaining a license to this product for purposes other than research, contact RheoGene, Inc. by calling Dr. Camille Jolly-Tornetta, Director of Intellectual Property (610-650-8734, ext. 104).
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