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pTO_CHMP4B_LAP
(Plasmid #101850)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 101850 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pTO_LAP_N-DEST
  • Backbone size w/o insert (bp) 6100
  • Total vector size (bp) 6772
  • Modifications to backbone
    Cloning procedure: 1. Starting vector: pcDNA5/FRT/TO (Invitrogen) 2. Cloning of LAP tag with Gateway cloning sites: starting point mCherry_N_dest 2.1. inserted a point mutation in XbaI site to remove Dam methylation site so it can be used for cloning (from GATC to GAAG) -> pmCherry_N_dest 2.2.exchanged mCherry with GFP using KpnI and XbaI --> pLAP_N_dest 3. Inserted LAP tag and Gateway cloning site from pLAP_N_dest into pcDNA5/FRT/TO -> pTO_LAP_N-DEST
  • Vector type
    Mammalian Expression, Bacterial Expression ; Gateway cloning, Flp-FRT recombination, Tet-inducible expression
  • Selectable markers
    Hygromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Charged Multivesicular Body Protein 4B
  • Alt name
    Charged Multivesicular Body Protein 4B
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    672
  • GenBank ID
    NM_176812.4
  • Entrez Gene
    CHMP4B (a.k.a. C20orf178, CHMP4A, CTPP3, CTRCT31, SNF7, SNF7-2, Shax1, VPS32B, Vps32-2, dJ553F4.4)
  • Promoter CMV
  • Tag / Fusion Protein
    • Localization and Affinity Purification (LAP) tag (C terminal on backbone)

Cloning Information

  • Cloning method Gateway Cloning
  • 5′ sequencing primer TACGGTGGGAGGTCTAT
  • 3′ sequencing primer TAGAAGGCACAGTCGAGG
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Part of the plasmid was derived from pmCherry_N_dest from the Helenius Lab (ETH Zuerich)

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pTO_CHMP4B_LAP was a gift from Daniel Gerlich (Addgene plasmid # 101850 ; http://n2t.net/addgene:101850 ; RRID:Addgene_101850)

  • For your References section:

    Dynamic subunit turnover in ESCRT-III assemblies is regulated by Vps4 to mediate membrane remodelling during cytokinesis. Mierzwa BE, Chiaruttini N, Redondo-Morata L, von Filseck JM, Konig J, Larios J, Poser I, Muller-Reichert T, Scheuring S, Roux A, Gerlich DW. Nat Cell Biol. 2017 Jul;19(7):787-798. doi: 10.1038/ncb3559. Epub 2017 Jun 12. 10.1038/ncb3559 PubMed 28604678
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