pALuWBB
(Plasmid #183202)

• Purpose: dsRed marked transgene for Crispr co-conversion with w+ gRNA and drop in cloning sites for 2 more gRNA
• Depositing Lab: Mitch McVey
• Publication: Drosophila Crispr co-conversion (unpublished)

Backbone

• Vector backbone: pCFD4
• Backbone manufacturer: Perrimon lab modified in our lab
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 5853
• Modifications to backbone: dsRed added as HindIII XhoI fragment linker between initial NsiI and XbaI site added
• Vector type: Insect Expression, CRISPR ; Recombines at attp landing site in fly genome

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Growth at 37C increases yield of plasmid.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: w+ gRNA and scaffold followed by 2 gRNA with BbsI and BsaI cloning sites
• gRNA/shRNA sequence: white gene and BbsI and BsaI sites for 2 other gRNA
• Species: D. melanogaster (fly)
• Promoter: U6-3

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: U6-3
• 3′ sequencing primer: M13 Rev

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry