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URL: http://www.addgene.org/42941

Proper Citation: RRID:Addgene_42941

Insert Name: Negative control Functionally Dead Human L1 element

Organism: Homo sapiens

Bacterial Resistance: Ampicillin

Defining Citation: PMID:16507671

Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:10410; Vector Backbone:pCEP4; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin

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. The A250V mutation found during Addgene QC is conservative and should not affect function.

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