pPL001
(Plasmid
#184750)
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PurposeBig-IN positive control payload encoding EF1a-driven GFP-T2A-BSD.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 184750 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepLM1110_LEU2
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Backbone manufacturerMaurano and Boeke labs, The Center for Synthetic Regulatory Genomics, NYU Langone Health
- Backbone size w/o insert (bp) 9932
- Total vector size (bp) 12551
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Vector typeMammalian Expression, Mouse Targeting, Cre/Lox, Synthetic Biology ; Recombinase-mediated cassette exchange (RMCE)
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Selectable markersBlasticidin
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature30°C
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Growth Strain(s)TransforMax EPI300
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Growth instructionsCopy number induction is recommended for high yield preps. See Lucigen’s growth and copy number induction protocol for TransforMax EPI300 cells
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namepEF1a-eGFP-T2A-BSD-bGHpA
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Alt nameHuman EF1a promoter driven CDS containing eGFP, T2A peptide and a blasticidin resistance gene. Bovine Growth Hormone Terminator
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SpeciesH. sapiens (human), B. taurus (bovine), Synthetic
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Insert Size (bp)2619
- Promoter EF1a
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer TGATCTCGCCCCGAGAACTG
- 3′ sequencing primer TCTATACCCGGGATCCCCAC (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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:
pPL001 was a gift from Jef Boeke (Addgene plasmid # 184750 ; http://n2t.net/addgene:184750 ; RRID:Addgene_184750) -
For your References section:
A versatile platform for locus-scale genome rewriting and verification. Brosh R, Laurent JM, Ordonez R, Huang E, Hogan MS, Hitchcock AM, Mitchell LA, Pinglay S, Cadley JA, Luther RD, Truong DM, Boeke JD, Maurano MT. Proc Natl Acad Sci U S A. 2021 Mar 9;118(10). pii: 2023952118. doi: 10.1073/pnas.2023952118. 10.1073/pnas.2023952118 PubMed 33649239