Phyco69
(Plasmid #255424)

• Purpose: Phyco69 contains a PLK region where any gene of interest can be inserted and get linked, through a short viral self-cleaving peptide (FMVD2A) to the 5` end of the APHVIII from S. rimosus
• Depositing Lab: Rosa León
• Publication: Molina-Marquez et al Int J Mol Sci. 2020 Jan 22;21(3):718. doi: 10.3390/ijms21030718.

Backbone

• Vector backbone: pBluscript II SK(+)
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 4041
• Total vector size (bp): 4842
• Modifications to backbone: Plasmid Phyco69 was constructed by including in front of the APHVIII gene a synthetic DNA fragment consisting of the final nucleotides of the promoter region of RbcS2, followed by the first intron of RbcS2, the 7 histidines, the FMDV-2A encoding sequences, and an artificial polylinker. This DNA fragment was inserted between the HindIII and BstBI restriction sites of pSI104, as described in (Molina-Márquez et al., 2020) which is a modification of pSI103 (Sizova et al 2001)
• Vector type: Plant Expression ; microalga expression vector
• Selectable markers: paromomycin

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: APHVIII
• Species: Streptomyces rimosus
• Insert Size (bp): 801
• Promoter: HSP70A-RBCS2
• Tags / Fusion Proteins:
  • 7His (N terminal on backbone)
  • MCS (N terminal on backbone)
  • FMDV2A (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BstBI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: T7 promoter taatacgactcactatagg
• 3′ sequencing primer: T3 promoter cctttagtgagggttaatt

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Phyco69 is based plasmid pSI103 (Sizova et al 2001) Sizova, I.; Fuhrmann, M.; Hegemann, P. A Streptomyces rimosus aphVIII gene coding for a new type phosphotransferase provides stable antibiotic resistance to Chlamydomonas reinhardtii. Gene 2001, 277, 221–229

Terms and Licenses

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