pAW-EGFP-4xlet-7 (pAW-AS27)
(Plasmid #40834)

• Purpose: miRNA sensor used in Drosophila cell lines
• Depositing Lab: Phillip Zamore
• Publication: Ameres et al Science. 2010 Jun 18;328(5985):1534-9.

Backbone

• Vector backbone: pAW
• Backbone manufacturer: Murhpy Lab, Carnegie Institution for Science
• Backbone size w/o insert (bp): 7233
• Vector type: Insect Expression ; miRNA reporter

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: Four target sites perfectly complementary to let-7
• Species: D. melanogaster (fly)
• Promoter: Actin5C
• Tag / Fusion Protein:
  • EGFP (N terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: EGFP-C

Terms and Licenses

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

Depositor Comments

The let-7 complimentary sequence was inserted in the XbaI site in the 3'UTR of EGFP using the following oligos:
s: CTA GAA CTA TAC AAC CTA CTA CCT CAT CTA GAA CTA TAC AAC CTA CTA CCT CAT
as: CTA GAT GAG GTA GTA GGT TGT ATA GTT CTA GAT GAG GTA GTA GGT TGT ATA GTT
This expression cassette was then sub-cloned into the Gateway vector pAW using GATEWAY cloning.