pRS403-PGAL1-hpSC_URA3
(Plasmid #22317)

• Depositing Lab: David Bartel
• Publication: Drinnenberg et al Science. 2009 Sep 10. ():.

Backbone

• Vector backbone: pRS403
• Backbone size w/o insert (bp): 4456
• Vector type: Yeast Expression ; Integrating plasmid
• Selectable markers: HIS3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): SURE
• Growth instructions: Provided in SURE cells (Stratagene)
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: URA hairpin
• gRNA/shRNA sequence: see View Sequence link below

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: T3
• 3′ sequencing primer: CYC1

Terms and Licenses

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

Depositor Comments

339bp of URA3 sequence from pRS406 was cloned into pYES2.1 GFP weak silencing construct in the sense orientation between PvuII and XhoI (replacing the GFP) and in antisense between EcoRI and XbaI sites. The expression cassette containing GAL1 promoter, URA3 silencing construct and CYC1 terminator was moved into pRS403.

The URA3 insert sequence can be found by clicking on the "View sequence" link. The rad9 intron loop is shown in upper case letters.

Please note that Addgene's sequencing results found a single nucleotide mismatch in the URA3 sequence at bp#3470 when compared to the full plasmid sequence provided by the depositing laboratory. This mismatch is not a concern for the function of the plasmid.