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URL: http://www.addgene.org/51288
Proper Citation: RRID:Addgene_51288
Insert Name: L1
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:21572950
Vector Backbone Description: Backbone Marker:Stratagene; Vector Backbone:pBluescript II; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: L1 is tagged with neoI indicator cassette containing an inverted neomycin resistance gene. The SV40 promoter (SV40p) drives transcription of the neomycin gene which is disrupted by an intron in the opposite orientation that renders it non-functional. The intron is only spliced out from tagged L1 transcripts generated by the CMV promoter. When retrotransposition of the spliced L1 RNA occurs, the new insert will contain a functional neo gene that is expressed from the SV40 promoter. The L1 open reading frames were codon optimized using Primo Optimum 3.4 (http://www.changbioscience.com/primo/primoo.html), which makes synonymous changes to optimal codons within the nucleotide sequence. A 63 bp inter-ORF with 92% sequence identity to L1RP was utilized for all constructs. Sequence changes to this region were generated to introduce restriction enzyme sites used for plasmid construction. The optimized nucleotide sequence is 74.9% identical to the sequence of L1RP.
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Source: Addgene
