pNUT N6His K206E/K534A hTFNG
(Plasmid #70133)

• Purpose: Expresses N-His tagged nonglycosylated human serum transferrin unable to release iron from the N-lobe or the C-lobe
• Depositing Labs: Ross MacGillivray, Anne Mason
• Publication: Halbrooks et al Biochemistry. 2003 Apr 8;42(13):3701-7.

Backbone

• Vector backbone: pNUT
• Backbone size w/o insert (bp): 7400
• Total vector size (bp): 9400
• 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: N6His K206e/K534A hTFNG
• Alt name: Mutant hTF double locked
• Species: H. sapiens (human)
• Insert Size (bp): 2136
• Mutation: changed Lys206 to a glutamic acid and Lys534 to an alanine,Asn413Asp and 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

This plasmid to express this double mutant of hTF was prepared in the laboratory of Dr. Ross T.A. MacGillivray using funding from my NIH grant. The plasmid was sent to Vermont for transfection, expression. purification and characterization of the resulting protein