pCI-Neo-p53mut
(Plasmid #99239)

• Purpose: Mutant p53 encoded by AS-30D rat hepatoma cell line
• Depositing Lab: Peter Pedersen
• Publication: Mathupala et al J Biol Chem. 1997 Sep 5;272(36):22776-80.

Backbone

• Vector backbone: pCI-Neo
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 5472
• Total vector size (bp): 6642
• Modifications to backbone: p53 cDNA inserted in MCS of vector between EcoRI and XbaI sites. A Kozak sequence was for enhanced translation initiation.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: p53
• Alt name: TP53
• Species: R. norvegicus (rat)
• Insert Size (bp): 1170
• Mutation: Silent mutations in translated protein at Gly (103) Ser and Glu (256) Gly.
• GenBank ID: U90328
• Entrez Gene: Tp53 (a.k.a. Trp53, p53)
• Promoter: CMV immediate/early enhancer/promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcorRI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: TAATACGACTCACTATAG
• 3′ sequencing primer: ATTAACCCTCACTAAAGGG

Terms and Licenses

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

Depositor Comments

Addgene's QC result finds an additional K318E mutation, which the depositor notes should have no functional consequence as it is away from the dimerization and DNA binding motifs