CDK1as_T2A_puromycin
(Plasmid #118595)

• Purpose: One of three plasmids required to create CDK1as human cells by single transfection (One shot system). CDK1as cDNA and puromycin resistance cassette flanked by Sleeping Beauty transposon.
• Depositing Lab: William Earnshaw
• Publication: Saldivar et al Science. 2018 Aug 24;361(6404):806-810. doi: 10.1126/science.aap9346.

Backbone

• Vector backbone: pT2/SVNeo, a gift from Perry Hackett (Addgene 26553 – Cui et al., 2002)
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: CDK1
• Species: X. laevis (frog)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NruI (destroyed during cloning)
• 3′ cloning site: ApaI (destroyed during cloning)
• 5′ sequencing primer: T7

Resource Information

• Supplemental Documents:
  • One shot method to create CDK1analogue-sensitive cell lines in human cells

Terms and Licenses

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

Depositor Comments

Please also cite the following publication:
Hochegger H, Dejsuphong D, Sonoda E, Saberi A, Rajendra E, Kirk J, Hunt T, Takeda S. An essential role for Cdk1 in S phase control is revealed via chemical genetics in vertebrate cells. J Cell Biol. 2007 Jul 16;178(2):257-68. PubMed PMID: 17635936; PubMed Central PMCID: PMC2064445.