pLVE-FH-IN
(Plasmid
#70071)
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Purpose(Empty Backbone) 2nd gen lentiviral vector. Cre-removable and tet/dox controllable expression of FLAG-HA-tagged transgene in mammalian cells
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Depositing Lab
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 70071 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonepLV-tTRKRAB-red
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Backbone manufacturerDr. Didier Trono, Addgene #12250
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Modifications to backboneadded FLAG-HA-IRES-neoR
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Vector typeMammalian Expression, Lentiviral, Cre/Lox
- Promoter EF1alpha
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Cloning Information
- Cloning method Restriction Enzyme
- 5′ sequencing primer EF-1a-F
- 3′ sequencing primer pCDH-rev (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
The DNA sequences of the IRES and neomycin resistant gene (neoR) were PCR amplified from pLV-tRKRAB-red (Dr. Didier Trono, Addgene plasmid 12250) and pEGFP-N1 (Clontech), respectively. 3' UTR of mouse Eef1a1 (NM_010106.2) was obtained from mouse embryonic stem cells by RT-PCR. Then, a BamHI-IRES-neoR (IN)-EcoRI-3'UTR of Eef1a1 (F)-Kpn1 fragment was synthesized by PCR flanked with PspXI-FLAG-HA(FH)-BamHI at the 5' end. Then the PspXI-FH-BamHI-IRES-neoR-EcoRI-E-KpnI fragment was subcloned into PspXI and KpnI sites of pLV-tTRKRAB-red to create pLVE-FH-IN.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pLVE-FH-IN was a gift from Thomas Tuschl (Addgene plasmid # 70071 ; http://n2t.net/addgene:70071 ; RRID:Addgene_70071) -
For your References section:
DND1 maintains germline stem cells via recruitment of the CCR4-NOT complex to target mRNAs. Yamaji M, Jishage M, Meyer C, Suryawanshi H, Der E, Yamaji M, Garzia A, Morozov P, Manickavel S, McFarland HL, Roeder RG, Hafner M, Tuschl T. Nature. 2017 Mar 23;543(7646):568-572. doi: 10.1038/nature21690. Epub 2017 Mar 15. 10.1038/nature21690 PubMed 28297718
