(829) SB Tet-ON dCasphi-[myc-ZIM3-HA] U6-gStop
(Plasmid #195622)

• Purpose: Sleeping Beauty Tet-ON plasmid for induced expression of dCasphi-ZIM3 CRISPRi construct
• Depositing Lab: Tonia Rex
• Publication: Casphi-ZIM3 CRISPRi plasmids (unpublished)

Backbone

• Vector backbone: Sleeping Beauty SBtet-Neo
• Backbone manufacturer: Addgene #60509
• Backbone size w/o insert (bp): 4600
• Total vector size (bp): 9033
• Modifications to backbone: Replaced neomycin-resistance gene with hygromycin-resistance gene that includes silent mutations for cloning purposes.
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: 34d
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: dCasphi
• Alt name: dCasJ2
• Species: Bacteriophage
• Insert Size (bp): 2300
• Mutation: P749A and P753A
• Promoter: Tight TRE promoter; Tet-ON (doxycycline)
• Tags / Fusion Proteins:
  • C-Myc (NLS) (C terminal on insert)
  • ZIM3 (C terminal on insert)
  • HA tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR1 (not destroyed)
• 3′ cloning site: Kpn1 (not destroyed)
• 5′ sequencing primer: SB-TRE-S
• 3′ sequencing primer: U6-Seq-A

Resource Information

• A portion of this plasmid was derived from a plasmid made by: IDT gBlock

Terms and Licenses

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