pmalc4x_E323K_TEV
(Plasmid #47699)

• Purpose: Expresses MBP myocilin olfactomedin domain fusion protein with a TEV protease recognition sequence and the mutation E323K
• Depositing Lab: Raquel Lieberman
• Publication: Burns et al Biochemistry. 2011 Jul 5;50(26):5824-33. doi: 10.1021/bi200231x. Epub 2011 Jun 9.

Backbone

• Vector backbone: pmalC4X
• Backbone manufacturer: NEB
• Backbone size w/o insert (bp): 6645
• Total vector size (bp): 7458
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Maltose Binding Protein and Olfactomedin-like domain with TEV cleavage site
• Alt name: MBP
• Alt name: OLF
• Species: H. sapiens (human)
• Insert Size (bp): 837
• Mutation: changed glutamic acid 323 to lysine
• Entrez Gene: MYOC (a.k.a. GLC1A, GPOA, JOAG, JOAG1, TIGR)
• Promoter: Ptac
• Tag / Fusion Protein:
  • MBP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: MBP-F
• 3′ sequencing primer: M13-F20; M13_pUC_Fwd

Terms and Licenses

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

Depositor Comments

The Lieberman Lab recommends E.coli Rosetta Gami 2 competent cells for protein expression.