pU6-neo-TET
(Plasmid #51286)

• Purpose: expresses U6 RNA, using its own pol III enhancer sequence and tagged with the self splicing intron neo cassette
• Depositing Lab: Astrid Roy-Engel
• Publication: Kroutter et al PLoS Genet. 2009 Apr;5(4):e1000458. doi: 10.1371/journal.pgen.1000458. Epub 2009 Apr 24.

Backbone

• Vector backbone: pAluYa5-neo-TET
• Backbone manufacturer: Addgene plasmid 51283
• 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: U6
• Alt name: HUMUG6
• Species: H. sapiens (human)
• Insert Size (bp): 512
• Entrez Gene: RNU6-1 (a.k.a. RNU6, RNU6A, U6, U6-1)
• Promoter: U6 upstream enhancer sequence
• Tag / Fusion Protein:
  • neoTET cassette with a tetrahymena self-splicing intron interrupting the neo gene (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: AatI (not destroyed)
• 5′ sequencing primer: M13-pUC-Fwd
• 3′ sequencing primer: SV40pro-F

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

U6 is tagged with the self splicing intron neo cassette such that G418 resistance will be obtained only if retrotransposition occurs.

Please note that the cassette is in the reverse orientation within the MCS relative to the full sequence--This has no functional consequence.