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MLM3743
(Plasmid #49962)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 49962 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pJFA274
  • Backbone manufacturer
    Joung lab (Addgene plasmid #49239)
  • Vector type
    TALEN

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    3x Flag Tet1CDmut HB-5
  • Alt name
    HB-5
  • Alt name
    human beta-globin (HBB) site #5
  • Species
    H. sapiens (human)
  • Mutation
    catalytic domain of TET1 inactive (H1671Y and D1673A mutations)
  • Entrez Gene
    HBB (a.k.a. CD113t-C, ECYT6, beta-globin)
  • Promoter EF1a
  • Tag / Fusion Protein
    • 3x FLAG (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (unknown if destroyed)
  • 3′ cloning site BsmBI (unknown if destroyed)
  • 5′ sequencing primer EF1a-F 5'-TCAAGCCTCAGACAGTGGTTC-3'
  • 3′ sequencing primer OJA199 5'-CACCATCACAGCAGTTGG-3'
    • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

TALE target site is: TATGCTTACCAAGCTGTGAT. Note that due to the TET1 mutations, this plasmid neither demethylated the proximal CpGs nor activated HBB gene expression.

See Supplementary Figure 13 of associated publication for amino acid sequences of backbone and TALE repeat array.

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:

    MLM3743 was a gift from Keith Joung (Addgene plasmid # 49962 ; http://n2t.net/addgene:49962 ; RRID:Addgene_49962)

  • For your References section:

    Targeted DNA demethylation and activation of endogenous genes using programmable TALE-TET1 fusion proteins. Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai SQ, Ho QH, Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson MF, Joung JK. Nat Biotechnol. 2013 Oct 9. doi: 10.1038/nbt.2726. 10.1038/nbt.2726 PubMed 24108092
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