pcDNA5 FRT TO myc Separase delta 55
(Plasmid #59824)

• Purpose: Allows the integration of myc Separase delta 55 in the genome and Tet-inducible expression.
• Depositing Lab: Stephen Taylor
• Publication: Holland et al J Biol Chem. 2007 Aug 24;282(34):24623-32. Epub 2007 Jun 29.

Backbone

• Vector backbone: pcDNA5/FRT/TO
• Backbone manufacturer: Invitrogen
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Separase
• Species: H. sapiens (human)
• Mutation: delta 55
• Entrez Gene: ESPL1 (a.k.a. ESP1, SEPA)
• Promoter: hybrid human cytomegalovirus (CMV)/TetO2 promoter
• Tag / Fusion Protein:
  • Myc (N terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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

Depositor Comments

To create separase delta 55, an N-terminal fragment (encoding amino acids 1-1418) was amplified and cloned as an XhoI-AgeI fragment. The C-terminal portion (amino acids 1474-2120) was then amplified and cloned as an AgeI-NotI fragment.