CMV Flag PAI-RBP
(Plasmid #45452)

• 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: pFlag CMV
• Backbone manufacturer: Sigma
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 5900
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PAI-RBP
• Alt name: DKFZp564M2423
• Alt name: Plasminogen activator inhibitor RNA binding protein
• Species: H. sapiens (human)
• Insert Size (bp): 1200
• GenBank ID: AL080119
• Entrez Gene: SERBP1 (a.k.a. CGI-55, CHD3IP, HABP4L, PAI-RBP1, PAIRBP1)
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: 5′-AAT GTC GTA ATA ACC CCG CCC CGT TGA CGC-3 Sigma
• 3′ sequencing primer: 5′-TAT TAG GAC AAG GCT GGT GGG CAC-3 Sigma

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 and then into CMV-Flag for mammalian expression.
Subsequently, we discovered isoforms of the protein with or without 6 and/or 15 amino acid insertions. These isoforms are now subcloned into pCEFL.
Additional reference: Heaton, JH et al 2003 Thromb Haemost 89; 959-966.