pAW-EGFP-2xmiR-1 (pAW-AS-21)
(Plasmid #40832)

• 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: Two target sites perfectly complementary to miR-1
• 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 miR-1 complimentary sequence was inserted in the XbaI site in the 3'UTR of EGFP using the following oligos:
s: CTA GAC TCC ATA CTT CTT TAC ATT CCA TCT AGA CTC CAT ACT TCT TTA CAT TCC AT
as: CTA GAT GGA ATG TAA AGA AGT ATG GAG TCT AGA TGG AAT GTA AAG AAG TAT GGA GT
This expression cassette was then sub-cloned into the Gateway vector pAW using GATEWAY cloning.