pLD564 human L1 ORF2p-3xFlag (L1RP, CMV promoter) in pCEP4 Puro
(Plasmid
#213030)
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PurposeExpresses full length human LINE-1 in human cells (native L1RP sequence), with a C-terminal 3xFlag tag on ORF2
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 213030 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCEP4 Puro
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Backbone manufacturerInvitrogen (modified)
- Backbone size w/o insert (bp) 9994
- Total vector size (bp) 16226
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Modifications to backboneReplacement of stock hygromycin with puromycin
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Vector typeMammalian Expression
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Selectable markersPuromycin
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameHuman LINE-1 (L1RP Sequence) with 5' UTR and ORF2-3xFlag
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SpeciesH. sapiens (human)
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Insert Size (bp)6232
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GenBank IDAF148856.1
- Promoter CMV
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Tag
/ Fusion Protein
- 3xFlag (ORF2p) (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site PmeI (destroyed during cloning)
- 5′ sequencing primer CMV_Forward
- 3′ sequencing primer SV40pA-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.
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:
pLD564 human L1 ORF2p-3xFlag (L1RP, CMV promoter) in pCEP4 Puro was a gift from Martin S. Taylor (Addgene plasmid # 213030 ; http://n2t.net/addgene:213030 ; RRID:Addgene_213030) -
For your References section:
Structures, functions and adaptations of the human LINE-1 ORF2 protein. Baldwin ET, van Eeuwen T, Hoyos D, Zalevsky A, Tchesnokov EP, Sanchez R, Miller BD, Di Stefano LH, Ruiz FX, Hancock M, Isik E, Mendez-Dorantes C, Walpole T, Nichols C, Wan P, Riento K, Halls-Kass R, Augustin M, Lammens A, Jestel A, Upla P, Xibinaku K, Congreve S, Hennink M, Rogala KB, Schneider AM, Fairman JE, Christensen SM, Desrosiers B, Bisacchi GS, Saunders OL, Hafeez N, Miao W, Kapeller R, Zaller DM, Sali A, Weichenrieder O, Burns KH, Gotte M, Rout MP, Arnold E, Greenbaum BD, Romero DL, LaCava J, Taylor MS. Nature. 2024 Feb;626(7997):194-206. doi: 10.1038/s41586-023-06947-z. Epub 2023 Dec 14. 10.1038/s41586-023-06947-z PubMed 38096902