pET-15b PAI-RBP
(Plasmid #45390)

• Depositing Lab: Thomas Gelehrter
• Publication: Heaton et al J Biol Chem. 2001 Feb 2;276(5):3341-7. Epub 2000 Sep 22.

Backbone

• Vector backbone: pET15b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5700
• Total vector size (bp): 6900
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use AG1 or BL21(DE3)pLysS E. coli for protein overexpression.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PAI-RBP
• Alt name: DKFZp564M2423
• Species: H. sapiens (human)
• Insert Size (bp): 1170
• GenBank ID: AL080119
• Entrez Gene: SERBP1 (a.k.a. CGI-55, CHD3IP, HABP4L, PAI-RBP1, PAIRBP1)
• Tag / Fusion Protein:
  • His (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nde I (not destroyed)
• 3′ cloning site: BamH I (not destroyed)
• 5′ sequencing primer: 5'-TAATACGACTCACTATAGGG-3' Novagen
• 3′ sequencing primer: 5'-GCTAGTTATTGCTCAGCGG-3' Novagen

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Resource Center and Primary Database of the German Human Genome Project

Terms and Licenses

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

Depositor Comments

Our lab isolated the protein based on it's ability to bind to a region of the PAI-1 (plasminogen activator inhibitor type 1) 3'UTR that we had determined to be involved in the cAMP regulation of mRNA stability. A search of the database revealed that the cDNA for the protein had been cloned by the German Genome Project and dubbed hypothetical protein. They kindly provided us with the clone, which we subcloned into pET-15b for expression of the protein.
Subsequently, we discovered isoforms of the protein with or without 6 and/or 15 amino acid insertions. These variants are now subcloned into pCELF.