pLCRcala (p53)
(Plasmid #12136)

• Depositing Lab: James Sherley
• Publication: Sherley et al J Biol Chem. 1991 Dec 25. 266(36):24815-28.

Backbone

• Vector backbone: pLCR0
• Backbone size w/o insert (bp): 5010
• Vector type: Mammalian Expression

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
• Species: M. musculus (mouse)
• Insert Size (bp): 2334
• Entrez Gene: Trp53 (a.k.a. Tp53, bbl, bfy, bhy, p44, p53)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: SV40pro-F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Arnold J. Levine (Princeton)
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

This plasmid, containing
the wild type murine p53 cDNA under the control of the previously
described BPV LCR-SV40 hybrid promoter (Spaholz et al., 1985),
was constructed as follows. Plasmid p407-1 (Spaholz et al., 1985) was
digested with EcoRI and HindIII, and the 5010-base pair fragment
containing the LCR-SV40 hybrid promoter was isolated and filled in
with Klenow.
After the ligation of BamHI linkers to the ends of the fragment, regenerating the terminal EcoRI site, the fragment was digested with
BamHI and ligated to the 2334-base pair BamHI fragment from the p53 cDNA expression vector pll-4 (Tan et al., 1986). This BamHI
fragment contained the entire wild type murine p53 coding region, 50 base pairs of 5'-untranslated sequence, 287 base pairs of 3'-untranslated
sequence, and a 3'-located SV40 mRNA splice site and polyA signal.