pBS-L1PA1-CH-mneo
(Plasmid #51288)

• Purpose: contains the fully codon optimized ORF1 and ORF2 of the human L1RP
• Depositing Lab: Astrid Roy-Engel
• Publication: Wagstaff et al PLoS One. 2011 May 10;6(5):e19672. doi: 10.1371/journal.pone.0019672.

Backbone

• Vector backbone: pBluescript II
• Backbone manufacturer: Stratagene
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: L1
• Alt name: L1RE1
• Alt name: Line1
• Species: H. sapiens (human)
• Entrez Gene: L1RE1 (a.k.a. L1.2, LRE1)
• Promoter: CMV
• Tag / Fusion Protein:
  • mneoI indicator cassette containing an inverted neomycin resistance gene (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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/pri​moo.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.