pMAL-c2x-MT1
(Plasmid #32101)

• Depositing Labs: David DeRosier, Chris Mercogliano, Cristina Risco Ortiz
• Publication: Mercogliano et al J Struct Biol. 2007 Oct;160(1):70-82. Epub 2007 Jul 10.

Backbone

• Vector backbone: pMAL-c2x
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 6700
• Vector type: Bacterial Expression

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: Metallothionein
• Species: M. musculus (mouse)
• Entrez Gene: Mt1 (a.k.a. MT-I, Mt-1)
• Promoter: P-lac
• Tag / Fusion Protein:
  • MBP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XmnI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: MBP_F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The metallothionein (MT) gene was amplified from pET-3d-MT (a gift from the Winge laboratory, University of Utah), which contains the MT gene within the pET-3d vector (Novagen, Madison, WI).

Terms and Licenses

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

Depositor Comments

The metallothionein (MT) gene was amplified with the forward primer (5'-CTCGGGATCGAGGGAAGGATTTCAAGA TATACCATGGACCCC-3'), which codes for the MBP linker region containing an XmnI and NcoI site fused in frame to the MT gene, and the reverse primer (5'-GACT CTAGAGGATCCTAGGCACAGCACGTG-3' ), which contains the MT gene stop codon and a subsequent BamHI site. Both the PCR product and pMAL-c2x vector were digested with XmnI and BamHI and were later ligated together to generate the final plasmid. In addition to this plasmid Addgene offers versions with 2 copies (32102), 3 copies (32115) and 4 copies (32100) of MT