plp-strongSC_GFP
(Plasmid #22312)

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

Backbone

• Vector backbone: pYES2.1 modified
• Backbone size w/o insert (bp): 5857
• Vector type: Yeast Expression ; Integrating plasmid
• Selectable markers: URA3

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: GFP-rad9-GFP
• Insert Size (bp): 623

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PvuII (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: GAL1
• 3′ sequencing primer: CYC1

Terms and Licenses

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

Depositor Comments

A MCS containing XhoI and EcoRI sites was cloned between the PvuII and XbaI sites of pYES2.1. 275bp of GFP sequence was cloned in the sense orientation between PvuII and XhoI sites and in antisense orientation between EcoRI and XbaI sites. A 73bp sequence spanning intron 1 from S. pombe rad9 was added between XhoI and EcoRI sites.

To make this plasmid integrating, the 2micron and f1 origins were replaced (using NheI and SpeI sites with sequence from S. castellii sc633:288301-289016 (amplified from genomic DNA with 5'-AAAAGCTAGCGATCCCTTATCAAATATGGTAC and 5'-AAAAACTAGTGTAGAATCCAGAGAATAGAATC).