pEB1N-mApple-LZ-LOV2
(Plasmid
#197418)
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PurposeMammalian expression of the mApple-tagged N-terminal half of photo-inactivated π-EB1
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 197418 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepEGFP-C1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 3936
- Total vector size (bp) 5798
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Modifications to backboneremoved EGFP
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameMAPRE1
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Alt nameEB1
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SpeciesH. sapiens (human)
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Insert Size (bp)1857
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MutationEB1 aa 1-185; resistant to EB1 shRNA#3 (Addgene #37927)
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GenBank IDNM_012325.2
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Entrez GeneMAPRE1 (a.k.a. EB1)
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Tags
/ Fusion Proteins
- GCN4 leucine zipper (C terminal on insert)
- mApple (C terminal on insert)
- A. sativa phototropin 1 LOV2 domain (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer CMV forward
- 3′ sequencing primer SV40 reverse (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 byEB1 from L. Cassimeris (Addgene #17234) LOV2 from Klaus Hahn (Addgene #22027)
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
LOV2 domain starts with LAAA reported to stabilize Jα helix docking (Strickland et al. PMID: 22388287)
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:
pEB1N-mApple-LZ-LOV2 was a gift from Torsten Wittmann (Addgene plasmid # 197418 ; http://n2t.net/addgene:197418 ; RRID:Addgene_197418) -
For your References section:
Optogenetic EB1 inactivation shortens metaphase spindles by disrupting cortical force-producing interactions with astral microtubules. Dema A, van Haren J, Wittmann T. Curr Biol. 2022 Mar 14;32(5):1197-1205.e4. doi: 10.1016/j.cub.2022.01.017. Epub 2022 Jan 31. 10.1016/j.cub.2022.01.017 PubMed 35090591