pAcGP67A - murine TfR
(Plasmid #12392)

• Depositing Lab: Pamela Bjorkman
• Publication: Bjorkman lab plasmids (unpublished)

Backbone

• Vector backbone: pAcGP67A
• Backbone manufacturer: Pharmingen
• Backbone size w/o insert (bp): 9761
• Vector type: Insect 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: transferrin receptor
• Alt name: TfR
• Species: M. musculus (mouse)
• Insert Size (bp): 2000
• Mutation: See comments section
• GenBank ID: NM_011638
• Entrez Gene: Tfrc (a.k.a. 2610028K12Rik, CD71, E430033M20Rik, Mtvr1, TFR, TFR1, TR, Trfr, p90)
• Tags / Fusion Proteins:
  • leader peptide from baculovirus protein gp67 (N terminal on insert)
  • 6X-His tag (N terminal on insert)
  • Factor Xa cleavage site (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EagI (not destroyed)
• 5′ sequencing primer: 5' TTAA AAT gAT AAC CAT CTC gC 3'
• 3′ sequencing primer: 5' gAA gCg ggT CCA AgT TTC CCT g 3'

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Gerhard Rank, PhD Rotary Bone Marrow Research Laboratories Royal Melbourne Hospital Royal Pde Parkville 3050 VIC Australia
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The murine TfR sequence bp 367-2292 (of the ORF) was cloned into the pAcGP67A vector. The construct includes a short 3' UTR (past the stop codon at bp 2292). The TfR fragment was PCR amplified to introduce an EcoRI site at the 5' end and an EagI site at the 3' end. This was subsequently cloned into the corresponding EcoRI/EagI sites of pAcGP67A. The murine TfR was fused 3' to a gene segment encoding the hydrophobic leader peptide from the baculovirus protein gp67, a 6X-His tag, and a factor Xa cleavage site.