pRTC2-puro
(Plasmid #190567)

• Purpose: Expresses the full-length human L1 (L1.3) retrotransposon and an engineered neomycin retrotransposition reporter to monitor retrotransposition efficiency in cultured mammalian cells.
• Depositing Lab: Anthony Furano
• Publication: Cook et al Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4298-303. doi: 10.1073/pnas.1416869112. Epub 2015 Mar 23.

Backbone

• Vector backbone: pCEP4, modified
• Backbone manufacturer: Life Technologies
• Backbone size w/o insert (bp): 9790
• Total vector size (bp): 17610
• Modifications to backbone: The original hygromycin resistance gene was replaced with the CMV promoter-driven pac gene encoding puromycin N-acetyl-transferase; the CMV promoter driving the insert was replaced with the SV40 early promoter; for additional details, please see Supporting Information in Cook, Jones and Furano, 2015, and refer to the annotated SnapGene file.
• Vector type: Mammalian Expression
• Selectable markers: Puromycin ; note that the plasmid is selectable with puromycin, but the retrotransposition reporter is selectable with G418.

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: L1 (L1.3) with reporter cassette
• Species: H. sapiens (human)
• Insert Size (bp): 7820
• Promoter: SV40

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Multiple; see Cook, Jones, and Furano, 2015 (not destroyed)
• 3′ cloning site: Multiple; see Cook, Jones, and Furano, 2015 (not destroyed)
• 5′ sequencing primer: see map and sequence to design custom primers for sequencing region of interest
• 3′ sequencing primer: see map and sequence to design custom primers for sequencing region of interest

Resource Information

• Supplemental Documents:
  • pRTC2-puro

Terms and Licenses

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

Depositor Comments

For full origin and modification details, please see Supporting Information in Cook, Jones, and Furano, 2015, 10.1073/pnas.1416869112, PMID: 25831499.