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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 17620 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneEF.CMV.GFP
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Vector typeMammalian Expression, Lentiviral
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Selectable markersGFP
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)Stbl3
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Copy numberUnknown
Gene/Insert
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Gene/Insert namedelFasL
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Alt nameFas ligand
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SpeciesM. musculus (mouse)
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Insert Size (bp)800
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MutationNoncleavable mouse delFasL . Deleted the exon 2 region, which contains the metalloproteinase cleavage site.
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Entrez GeneFasl (a.k.a. APT1LG1, CD178, CD95-L, CD95L, Fas-L, Faslg, Tnfsf6, Tnlg1a, gld)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ sequencing primer EF-1a forward (Common Sequencing Primers)
Resource Information
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Addgene Notes
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 CMV promoter was inserted immediately upstream of GFP in the EF.GFP LV to make the intermediate LV, EF.V-CMV.GFP, with a unique EcoRV site available to insert a gene of interest under the control of the EF promoter. Noncleavable mouse delFasL cDNA was constructed using PCR cloning to delete the exon 2 region, which contains the metalloproteinase cleavage site. The delFasL gene was then cloned into the EcoRV site of EF.CMV.GFP to make the EF.delFasL-CMV.GFP LV.
Recombinant LVs were produced by transient transfection of the transducing vector into 293T cells with two packaging vectors: pMD.G, a plasmid expressing the VSV-G envelope gene, and pCMVDeltaR8.91, a plasmid expressing the HIV-1 gag/pol, tat, and rev genes.
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:
EF.delmFasL.CMV.GFP was a gift from Linzhao Cheng (Addgene plasmid # 17620 ; http://n2t.net/addgene:17620 ; RRID:Addgene_17620) -
For your References section:
Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells. Yu X, Zhan X, D'Costa J, Tanavde VM, Ye Z, Peng T, Malehorn MT, Yang X, Civin CI, Cheng L. Mol Ther. 2003 Jun . 7(6):827-38. 10.1016/S1525-0016(03)00104-7 PubMed 12788657