pNUT N6His K206E hTFNG
(Plasmid #70130)

• Purpose: Expresses N-His tagged nonglycosylated human serum transferrin unable to release iron from the N-lobe
• Depositing Labs: Ross MacGillivray, Anne Mason
• Publication: He et al Biochemistry. 1999 Jul 27;38(30):9704-11.

Backbone

• Vector backbone: pNUT
• Backbone size w/o insert (bp): 7400
• Total vector size (bp): 9540
• Vector type: Mammalian Expression
• Selectable markers: Methotrexate

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: mutated human serum transferrin
• Alt name: K206E or N-locked hTF
• Species: H. sapiens (human)
• Insert Size (bp): 2136
• Mutation: changed Lys206 to a glutamic acid, Asn413Asp, Asn611Asp
• GenBank ID: AF28814
• Entrez Gene: TF (a.k.a. HEL-S-71p, PRO1557, PRO2086, TFQTL1)
• Promoter: SV40
• Tag / Fusion Protein:
  • N-terminal signal peptide, 4 aa link, 6 His, Factor Xa cleavage site (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SmaI (not destroyed)
• 3′ cloning site: SmaI (not destroyed)
• 5′ sequencing primer: CCTTTGCGCCCGGACTCG
• 3′ sequencing primer: GTGGCAACTTCCAGGGCC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The laboratory of Dr. Ross T.A. MacGillivray at UBC in Vancouver Canada

Terms and Licenses

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

Depositor Comments

The MacGillivray lab constructed the plasmid with funds from my NIH grant . They sent it to my lab for transfection of BHK cells, expression, purification and characterization of the inserted mutant