pDA186
(Plasmid #74250)

• Purpose: Intermediate cloning vector, with MCS2 containing inducible unstable peptide driven by pGPD1 (UbiY 2xNLSs SZ3).
• Depositing Lab: Serge Pelet
• Publication: Aymoz et al Nat Commun. 2016 Apr 21;7:11304. doi: 10.1038/ncomms11304.

Backbone

• Vector backbone: pRS305
• Backbone size w/o insert (bp): 5053
• Total vector size (bp): 7088
• Modifications to backbone: The MCS was removed by PvuII digestion. The coding sequence is now flanked by AatII and SphI.
• Vector type: Bacterial Expression, Yeast Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Inducible peptide of the dPSTR
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 2036
• Promoter: pGPD1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AatII (not destroyed)
• 3′ cloning site: SphI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Resource Information

• Supplemental Documents:
  • Serial Cloner annotated sequence file
• A portion of this plasmid was derived from a plasmid made by: The SynZips were provided by the Keating lab.

Terms and Licenses

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

Depositor Comments

This intermediate cloning vector is used to change the measured promoter (here pGPD1), by cloning it between SacI (or AatII) and XbaI (or SpeI). The entire inducible peptide has to be inserted into a pSIV by cloning AatII / SphI (into pDA183 or pDA200 for instance).
For more information, see Aymoz et al. Nat. Comm. 2016.