pcDNA3.2 V5-DEST mir-1-1 reporter
(Plasmid #46646)

• Purpose: expression of query pri-miRNA
• Depositing Lab: David Bartel
• Publication: Auyeung et al Cell. 2013 Feb 14;152(4):844-58. doi: 10.1016/j.cell.2013.01.031.

Backbone

• Vector backbone: pcDNA3.2/V5-DEST
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 7711
• Vector type: Mammalian Expression, RNAi ; Expression of query pri-miRNA sequences in HEK293 cells
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mir-1-1 reporter
• Promoter: CMV
• Tag / Fusion Protein:
  • V5 (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: TK-pA-R

Resource Information

• Supplemental Documents:
  • Annotated GenBank file

Terms and Licenses

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

Depositor Comments

Query pri-miRNA sequences are to be recombined into this plasmid using Gateway recombination. The human pri-mir-1-1 sequence is cloned downstream of the query pri-miRNA and is transcriptionally fused to the query pri-miRNA sequence. This plasmid was used for ectopic pri-miRNA expression in HEK293 cells