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pRT006.3 L1 dual-luciferase AI retrotransposition reporter
(Plasmid #213031)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 213031 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pCEP Puro
  • Backbone manufacturer
    Genscript / Invitrogen
  • Backbone size w/o insert (bp) 8859
  • Total vector size (bp) 18643
  • Vector type
    Mammalian Expression, Luciferase
  • Selectable markers
    Puromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Tet-On Human LINE-1 (ORFeus-Hs) Firefly + Renilla Luciferase Antisense Intron dual retrotransposition marker
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    9784
  • Promoter pTRE bidirectional tet-on
  • Tag / Fusion Protein
    • Firefly luciferase antisense intron (3' UTR) (C terminal on backbone)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer bidir_pTRE_Fwd (CGCCACCTCGACATACTCG)
  • 3′ sequencing primer SV40pA-Rev
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The construct is based on the synthetic L1 ORFeus-Hs sequence generated in the lab of Jef Boeke (see W. An et al., Mobile DNA 2, 2011 https://doi.org/10.1186/1759-8753-2-2)

For integration with Sleeping Beauty such as SB100x (Izsvak lab, Addgene #34879). This vector is similar to pYX056 from An lab, see also:
Xie, Y. et al. Cell division promotes efficient retrotransposition in a stable L1 reporter cell line. Mob DNA 4, 10 (2013). https://doi.org/10.1186/1759-8753-4-10
and
Xie, Y., Rosser, J. M., Thompson, T. L., Boeke, J. D. & An, W. Characterization of L1 retrotransposition with high-throughput dual-luciferase assays. Nucleic Acids Res 39, e16 (2011). https://doi.org/10.1093/nar/gkq1076

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:

    pRT006.3 L1 dual-luciferase AI retrotransposition reporter was a gift from Donna Romero & Martin S. Taylor (Addgene plasmid # 213031 ; http://n2t.net/addgene:213031 ; RRID:Addgene_213031)

  • For your References section:

    Structures, functions and adaptations of the human LINE-1 ORF2 protein. Baldwin ET, van Eeuwen T, Hoyos D, Zalevsky A, Tchesnokov EP, Sanchez R, Miller BD, Di Stefano LH, Ruiz FX, Hancock M, Isik E, Mendez-Dorantes C, Walpole T, Nichols C, Wan P, Riento K, Halls-Kass R, Augustin M, Lammens A, Jestel A, Upla P, Xibinaku K, Congreve S, Hennink M, Rogala KB, Schneider AM, Fairman JE, Christensen SM, Desrosiers B, Bisacchi GS, Saunders OL, Hafeez N, Miao W, Kapeller R, Zaller DM, Sali A, Weichenrieder O, Burns KH, Gotte M, Rout MP, Arnold E, Greenbaum BD, Romero DL, LaCava J, Taylor MS. Nature. 2024 Feb;626(7997):194-206. doi: 10.1038/s41586-023-06947-z. Epub 2023 Dec 14. 10.1038/s41586-023-06947-z PubMed 38096902
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