pAGE1.0
(Plasmid #165983)

• Purpose: Multi-host shuttle vector that can be transferred by conjugation and replicate in S. meliloti, E. coli, S. cerevisiae, and P. tricornutum. Contains S. meliloti pSymA origin and Spec resistance.
• Depositing Lab: Bogumil Karas
• Publication: Brumwell et al PLoS One. 2019 Jun 17;14(6):e0206781. doi: 10.1371/journal.pone.0206781. eCollection 2019.

Backbone

• Vector backbone: pPtGE31
• Backbone manufacturer: Slattery et al., 2018
• Backbone size w/o insert (bp): 9378
• Total vector size (bp): 18129
• Modifications to backbone: Addition of elements for S. meliloti replication and selection.
• Vector type: Synthetic Biology ; Bacterial and Yeast Cloning; Conjugation
• Selectable markers: HIS3 ; Chloramphenicol - E. coli, Spectinomycin - S. meliloti, Nourseothricin N-acetyltransferase - P. tricornutum

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): E. coli Epi300
• Growth instructions: Plasmid can be induced to high copy with arabinose.
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: RK2/RP4 origin of transfer
• Alt name: oriT
• Insert Size (bp): 771

Gene/Insert 2

• Gene/Insert name: repA2B2C2
• Species: Sinorhizobium meliloti
• Insert Size (bp): 4335

Gene/Insert 3

• Gene/Insert name: nourseothricin N-acetyl transferase
• Alt name: NAT
• Insert Size (bp): 570

Gene/Insert 4

• Gene/Insert name: spectinomycin/streptomycin resistance
• Alt name: aadA1
• Insert Size (bp): 2066

Resource Information

• Supplemental Documents:
  • pAGE1-0-sequenced.gb
• A portion of this plasmid was derived from a plasmid made by: (1) Edgell Lab (backbone was cloned from pPtGE30). Reference: Slattery, S. S., Diamond, A., Wang, H., Therrien, J. A., Lant, J. T., Jazey, T., ... & Edgell, D. R. (2018). An expanded plasmid-based genetic toolbox enables Cas9 genome editing and stable maintenance of synthetic pathways in Phaeodactylum tricornutum. ACS synthetic biology, 7(2), 328-338. (2) Frey and Krisch Lab (SpecR/StrepR resistance gene was cloned from the Ω cassette). Reference: Frey, J., & Krisch, H. M. (1985). Ω mutagenesis in gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species. Gene, 36(1-2), 143-150.

Terms and Licenses

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