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PurposeDual fluorescent reporter - constitutive expression of mVenus (515/528) localized in the nucleus, mCherry (587/610) untargeted and visualized in the cytoplasm, and Puromycin resistance
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 99278 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneMillerLab H2B-mVenus-IRES-mPlum, DNA 2.0 Order #62961, Gene 179575
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Backbone manufacturerDNA 2.0
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Modifications to backbonecut out mPlum using NheI and BamHI, and ligate in mCherry
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Vector typeLentiviral
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Selectable markersPuromycin
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberUnknown
Gene/Insert 1
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Gene/Insert nameH2B-mVenus-IRES-mCherry
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SpeciesH. sapiens (human)
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Insert Size (bp)2434
- Promoter EF1 alpha-human
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Tags
/ Fusion Proteins
- mVenus (N terminal on insert)
- H2B (N terminal on insert)
- mCherry (N terminal on insert)
- IRES (N terminal on insert)
Cloning Information for Gene/Insert 1
- Cloning method Unknown
- 5′ sequencing primer not sure
- 3′ sequencing primer not sure (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert namePuro
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Alt namePuromycin
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SpeciesM. musculus (mouse)
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Insert Size (bp)600
- Promoter PGK-mouse
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Tag
/ Fusion Protein
- PGK (N terminal on backbone)
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer not sure (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byH2B-Venus sequence from Anna-Katerina Hadjantonakis: Addgene plasmid # 60141 (https://www.addgene.org/browse/sequence/60141/) Nowotschin, S., Xenopoulos, P., Schrode, N., & Hadjantonakis, A.-K. (2013). A bright single-cell resolution live imaging reporter of Notch signaling in the mouse. BMC Developmental Biology, 13, 15. doi:10.1186/1471-213X-13-15
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Article 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:
hEF1α-H2B-mVenus-IRES-mCherry PGK-Puromycin was a gift from Jordan Miller (Addgene plasmid # 99278 ; http://n2t.net/addgene:99278 ; RRID:Addgene_99278) -
For your References section:
Tubulogenesis of co-cultured human iPS-derived endothelial cells and human mesenchymal stem cells in fibrin and gelatin methacrylate gels. Calderon GA, Thai P, Hsu CW, Grigoryan B, Gibson SM, Dickinson ME, Miller JS. Biomater Sci. 2017 Jun 29. doi: 10.1039/c7bm00223h. 10.1039/c7bm00223h PubMed 28661522