phBG-P53DD
(Plasmid #149707)

• Purpose: In vitro transcription of dominant negative form of P53 mRNA for RNA transfection into mammalian cells
• Depositing Lab: Erika Sasaki
• Publication: Watanabe et al Genes Cells. 2019 Jul;24(7):473-484. doi: 10.1111/gtc.12702. Epub 2019 Jun 17.

Backbone

• Vector backbone: pCR4 Blut TOPO
• Backbone size w/o insert (bp): 3947
• Total vector size (bp): 4653
• Modifications to backbone: Alphan globin 5'UTR is added at the downstream of T7 promoter Tandem human beta globin 3' UTR is added
• Vector type: RNA transcription

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: Trp53
• Alt name: P53
• Species: M. musculus (mouse)
• Insert Size (bp): 387
• Mutation: deleted amino acid 14-301
• Entrez Gene: Trp53 (a.k.a. Tp53, bbl, bfy, bhy, p44, p53)
• Promoter: T7

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Not1 (not destroyed)
• 3′ cloning site: Asc (not destroyed)
• 5′ sequencing primer: M13 Forward
• 3′ sequencing primer: M13 Revrse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pMXs-p53DD (Addgene#22729)

Terms and Licenses

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

Depositor Comments

This plasmid contains a ccdB marker in the backbone which appears to be nonfunctional/untranscribed under routine laboratory propagation in E. coli. If problems during routine propagation arise, the use of a ccdB tolerant strain is suggested.