pGS1021
(Plasmid #59575)

• Purpose: Vector to create a cassette for homologous recombination in S. cerevisiae; adds a C-terminal Twin-Strep tag and a TRP1 selection marker
• Depositing Labs: Monika Golas, Bjoern Sander
• Publication: Rai et al Mol Biotechnol. 2014 Jun 27.

Backbone

• Vector backbone: pUC57
• Backbone manufacturer: Fermentas/Thermo
• Backbone size w/o insert (bp): 2710
• Total vector size (bp): 3824
• Vector type: E. coli cloning vector
• Selectable markers: TRP1

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Twin-Strep tag optimized for PCR amplification followed by TRP1 selection marker
• Species: Synthetic
• Insert Size (bp): 1171
• Promoter: None
• Tag / Fusion Protein:
  • Twin-Strep tag (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: SphI (not destroyed)
• 5′ sequencing primer: AGG GTT TTC CCA GTC ACG ACG TT
• 3′ sequencing primer: GAG CGG ATA ACA ATT TCA CAC AGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Twin-Strep tag sequence was amplified from a commercial plasmid pPRIBA101 (IBA, Goettingen, Germany) and mutated by site-directed mutagenesis to an improved sequence for PCR amplification of the tagging cassette. The encoded amino acids were not changed by this mutagenesis.

Terms and Licenses

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