pCI-Neo-HKIImut
(Plasmid #99240)

• Purpose: Type II Hexokinase expressed in AS-30D rat hepatoma cell line
• Depositing Lab: Peter Pedersen
• Publication: Mathupala et al J Biol Chem. 1997 Sep 5;272(36):22776-80.

Backbone

• Vector backbone: pCI-Neo
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 5472
• Total vector size (bp): 8220
• Modifications to backbone: HKII cDNA (cloned from rat AS-30D hepatoma) inserted in MCS of vector between EcoRI and XbaI sites. A Kozak sequence was for enhanced translation initiation.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Hexokinase II
• Alt name: HKII
• Alt name: Type II hexokinase
• Alt name: HK2
• Species: R. norvegicus (rat)
• Insert Size (bp): 2748
• Mutation: Four silent mutations; P (115)L, A (192) V, F (782) S, L (854) P
• GenBank ID: AF027179
• Entrez Gene: Hk2
• Promoter: CMV I.E.

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: TAATACGACTCACTATAG
• 3′ sequencing primer: ATTAACCCTCACTAAAGGG

Terms and Licenses

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

Depositor Comments

Addgene's QC result found 4 additional mutations (V192A, V378A, M555V, and E699G), that are unexpected; however, the depositor notes that they are away from the catalytic site (and the catalytic site is active).