pET MBP mRuby2 LIC cloning vector (MBP-mRuby2)
(Plasmid #86933)

• Purpose: (Empty Backbone) pET LIC cloning vector for Biotin-His6-MBP-N10-tev-yORF-mRuby2
• Depositing Lab: Chris Jeans
• Publication: C-terminal fluorescent fusion vectors for E. coli expression (unpublished)

Backbone

• Vector backbone: pET
• Backbone size (bp): 7267
• Modifications to backbone: mClover is from AddGene plasmid 49089.
• Vector type: Bacterial Expression
• Promoter: T7
• Tags / Fusion Proteins:
  • Biotin (N terminal on backbone)
  • His6 (N terminal on backbone)
  • MBP (N terminal on backbone)
  • Tev Recognition Sequence (N terminal on backbone)
  • mRuby2 (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: MBP F (5' ggtcgtcagactgtcgatgaagcc 3')
• 3′ sequencing primer: T7 Reverse

Resource Information

• Supplemental Documents:
  • MBP-mRuby2.gb
• A portion of this plasmid was derived from a plasmid made by: The sequence for mRuby2 came from Addgene #49089 (pcDNA3.1-Clover-mRuby2).

Terms and Licenses

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

Depositor Comments

This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. This plasmid can be used as a single-expression vector, and it is also compatible with our 2-series polycistronic destination vectors (2D, 2E, and 2Z), if co-expression with other genes is desired.

To clone into this vector, add LIC tags to the 5' end of your PCR primers.

Forward - 5' TACTTCCAATCCAATGCA 3'

Reverse - 5' CTCCCACTACCAATGCC 3'

Do NOT include a stop codon with your reverse primer.

Linearize the plasmid with SspI, then gel purify.

When digesting the DNA with T4 polymerase, use dGTP for the insert and dCTP for your linearized vector.

More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/