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Purpose(Empty Backbone) N-terminal 3xFLAG tag. Gateway destination vector for lentivirus production and stable transduction in mammalian cells.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 123223 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepLV-CMV-y/hNubI-tripleFLAG-linker-Gateway-PuroR
- Backbone size (bp) 9000
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Modifications to backboneReplacement of region between NheI and BstBI sites with 3xFLAG and gateway cassette.
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Vector typeMammalian Expression, Lentiviral ; Gateway destination
- Promoter CMV
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Selectable markersPuromycin
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Tag
/ Fusion Protein
- 3xFLAG (N terminal on backbone)
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol and Ampicillin, 25 & 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)ccdB Survival
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Growth instructionsChloramphenicol selection prevents recombination of unmodified backbone. However, expression clones generated using this construct are prone to recombination, and must be grown using a recombination deficient strain.
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Copy numberHigh Copy
Cloning Information
- Cloning method Gateway Cloning
- 5′ sequencing primer CMVF_pCDNA3
- 3′ sequencing primer cPPT-rev (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Articles Citing this Plasmid
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.
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:
pLVpuro-CMV-N-3xFLAG was a gift from Robin Ketteler (Addgene plasmid # 123223 ; http://n2t.net/addgene:123223 ; RRID:Addgene_123223) -
For your References section:
Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells. Agrotis A, Pengo N, Burden JJ, Ketteler R. Autophagy. 2019 Jan 21. doi: 10.1080/15548627.2019.1569925. 10.1080/15548627.2019.1569925 PubMed 30661429