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EF06R
(Plasmid #42940)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 42940 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCEP4
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 10410
  • Total vector size (bp) 23135
  • Modifications to backbone
    Blunt ligation of nt 1-1392 of pPur (BD/Clontech) into the NruI site of pCEP4 to insert PuroR
  • Vector type
    Mammalian Expression
  • Selectable markers
    Puromycin, Hygromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Human L1 element
  • Alt name
    LINE-1
  • Alt name
    Long Interspersed Element 1
  • Alt name
    L1RT
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    7904
  • GenBank ID
    M80343.1
  • Entrez Gene
    L1RE1 (a.k.a. L1.2, LRE1)
  • Promoter CMV MIE, L1 5'UTR
  • Tag / Fusion Protein
    • EGFP (opposite strand) (C terminal on insert)

Cloning Information

Resource Information

Depositor Comments

These plasmids consist of the L1RP element tagged with an enhanced green fluorescent protein (EGFP) cassette (pL1RP-EGFP) in a pCEP4 backbone (Invitrogen). The L1 element is driven by its 5′-UTR and an upstream CMV MIE promoter. L1-EGFP (EF06R, Addgene plasmid #42940) was derived from L1RP-EGFP by blunt ligation of 1–1392 nt of pPur (BD/Clontech) into the NruI site of pCEP4.

A negative control ‘dead’ L1-EGFP (EF05J, Addgene plasmid #42941) was similarly derived from pL1RP(JM111)-EGFP, which contains disabling mutations in ORF1.

An EN− plasmid (EF13E, Addgene plasmid #42942) was created by swapping 1927–3708 nt (Age I–Bcl I) from JM102D205A, which contains an endonuclease function-abrogating D205A point mutation in the EN domain, into EF06R.

A control plasmid (EF12J, Addgene plasmid #42943) was similarly derived from JM102. JM102 and JM102D205A are derived from L1.3, which differs from L1RP by 10 base changes in the region swapped.

I->T substitution at the end of ORF 2 should not affect plasmid function.

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:

    EF06R was a gift from Eline Luning Prak (Addgene plasmid # 42940 ; http://n2t.net/addgene:42940 ; RRID:Addgene_42940)
  • For your References section:

    Gamma radiation increases endonuclease-dependent L1 retrotransposition in a cultured cell assay. Farkash EA, Kao GD, Horman SR, Prak ET. Nucleic Acids Res. 2006 Feb 28;34(4):1196-204. Print 2006. 10.1093/nar/gkj522 PubMed 16507671