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pEM184_30x147bp_Widom601
(Plasmid #240634)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 240634 Standard format: Plasmid sent in bacteria as agar stab 1 $89
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Backbone

  • Vector backbone
    pMLS280
  • Backbone manufacturer
    Erik Jorgensen (Addgene plasmid #73738)
  • Backbone size w/o insert (bp) 2438
  • Total vector size (bp) 7202
  • Modifications to backbone
    A backbone SapI site in pMLS280 was changed to an NdeI site
  • Vector type
    DNA amplification

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    We routinely grow in NEB 5-alpha for amplifying large quantities from the sequenced plasmid stock. Due to the repetitive sequence, for propagation and distribution, NEB Stable would likely be better suited.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    30x 147-bp Widom 601
  • Species
    Synthetic
  • Insert Size (bp)
    4764

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site KpnI (not destroyed)
  • 3′ cloning site BamHI, BglII (destroyed during cloning)
  • 5′ sequencing primer T7 promoter, M13 forward
  • 3′ sequencing primer T3 promoter, M13 reverse
    • (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

Cloning carried out as in: Dyer PN, Edayathumangalam RS, White CL, Bao Y, Chakravarthy S, Muthurajan UM, Luger K. Reconstitution of nucleosome core particles from recombinant histones and DNA. Methods Enzymol. 2004;375:23-44. doi: 10.1016/s0076-6879(03)75002-2. PMID: 14870657.

DNA purification as in: Paintsil EA, Morrison EA. Preparation of Recombinant Histones and Widom 601 DNA for Reconstitution of Nucleosome Core Particles. Methods Mol Biol. 2023;2599:163-175. doi: 10.1007/978-1-0716-2847-8_12. PMID: 36427149.

Nucleosome core particle reconstitutions largely as in: Hammonds EF, Morrison EA. Nucleosome Core Particle Reconstitution with Recombinant Histones and Widom 601 DNA. Methods Mol Biol. 2023;2599:177-190. doi: 10.1007/978-1-0716-2847-8_13. PMID: 36427150. Exception: Suggest reconstitution molar ratios of 1:1:2 DNA:H3/H4-tetramer:H2A/H2B-dimer.

Please visit https://doi.org/10.1101/2025.04.09.647968 for bioRxiv preprint.

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:

    pEM184_30x147bp_Widom601 was a gift from Emma Morrison (Addgene plasmid # 240634 ; http://n2t.net/addgene:240634 ; RRID:Addgene_240634)

  • For your References section:

    Histone H3 tail charge patterns govern nucleosome condensate formation and dynamics. Hammonds EF, Singh A, Suresh KK, Yang S, Zahorodny SSM, Gupta R, Potoyan DA, Banerjee PR, Morrison EA. bioRxiv [Preprint]. 2025 Apr 10:2025.04.09.647968. doi: 10.1101/2025.04.09.647968. 10.1101/2025.04.09.647968 PubMed 40291647
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