pEN_TmiR_G13-L_G12-E
(Plasmid #25763)

• Purpose: Entry vector with TRE promoter driving both mouse G alpha 12 and G alpha 13 miR30-based shRNAs.
• Depositing Lab: Iain Fraser
• Publication: Shin et al Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.

Backbone

• Vector backbone: pENTR1A-Gent
• Backbone manufacturer: ATCC 10326362
• Backbone size w/o insert (bp): 4641
• Vector type: Entry vector

Growth in Bacteria

• Bacterial Resistance(s): Gentamicin, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Growth instructions: Top10
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: G alpha 13 and 12 miR-shRNA
• Species: M. musculus (mouse)
• Insert Size (bp): 22

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: attL1 (not destroyed)
• 3′ cloning site: attL2 (not destroyed)
• 5′ sequencing primer: pCEP-fwd
• 3′ sequencing primer: n/a

Resource Information

• A portion of this plasmid was derived from a plasmid made by: TRE promoter- David Anderson, Caltech; miR30- Greg Hannon, CSHL

Terms and Licenses

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

Depositor Comments

Entry vector with TRE-driven G alpha 12 and 13 miR-shRNA

G alpha 12 miR-shRNA: GCGACACCATCTTCGACAACAT

G alpha 13 miR-shRNA: CTGGGTGAGTCTGTAAAGTATT

There are 3 minor changes between depositor's sequence and Addgene's sequence - An extra T between the TRE promoter and first shRNA; a 17nt stretch between the shRNAs is different but will not affect function; and a single nt change mutates an EcoRI site just beyond the 2nd shRNA but will again not affect function.