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- MCS is downstream of 5 copies of GAL4 RE (5XRE) and a minimal promoter.
- When used in conjunction with pNEBR-R1 Regulator Plasmid, expression of a cloned gene is controlled by the addition of RSL1.
- Contains hygromicin resistance gene for the selection of stable cell lines.
- Does not contain any translation initiation signals. These must be part of the cloned insert DNA.
- Ampicillin resistance for propagation in E. coli.
- RheoSwitch Primer (NEB #S1280S) and T7 Minimal Promoter Primer (NEB #S1272S) can be used to sequence across the MCS.
Description:
pNEBR-X1Hygro Vector has 5 copies of the GAL4 RE (5XRE) upstream of a minimal TATA signal and multiple cloning site (MCS) and SV40 polyA signal. It is used to clone and express a gene of interest in mammalian cells as part of the RheoSwitch® Mammalian Inducible Expression System (NEB #E3000). pNEBR-X1Hygro contains a hygromycin resistance cassette that allows for the construction of stable cell lines expressing the gene of interest. When co-transfected with pNEBR-R1 Regulator Plasmid in mammalian cells, expression of the cloned gene can be controlled by addition of the RheoSwitch Ligand, RSL1 (NEB #E3301).
pNEBR-X1Hygro plasmid map. Only unique restriction enzyme sites are shown.
pNEBR-X1Hygro multiple cloning site (MCS). To minimize uninduced expression, a short sequence containing the TATA box separates the end of the 5XRE from the predicted transcription start. Only unique restriction enzyme sites are shown.
Source:
pNEBR-X1 Vector is isolated from an 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
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- RheoSwitch Mammalian Inducible Expression System Manual., unpublished observations.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.
- Verhaegent, M. and Christopoulos, T.K. (2002) Anal. Chem., 74, 4378-4385.
Companion Products
pNEBR-R1 Regulator Plasmid
pNEBR-X1GLuc Control Plasmid
RheoSwitch® Cell Line HEK293-A7
RheoSwitch® Cell Line NIH3T3-47
RheoSwitch® Ligand RSL1
RheoSwitch® Mammalian Inducible Expression System
TransPass™ D1 Transfection Reagent
TransPass™ D2 Transfection Reagent
Legal
Patents:
RheoGene: U.S. Patent No. 6,258, 603
RheoGene: U.S. Patent No. 5,514,578
RheoGene: U.S. Patent No. 6,245,531
RheoGene: U.S. Patent No. 5,530,082
Licenses/Patents/Disclaimers:
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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