pFGL1252_TetOFF(Hyg)
(Plasmid #118992)

• Purpose: TetOFF (promoter-less) cassette with Hygromycin selection
• Depositing Lab: Naweed Naqvi
• Publication: Yadav et al MBio. 2019 Jul 30;10(4). pii: mBio.01581-19. doi: 10.1128/mBio.01581-19.

Backbone

• Vector backbone: pFGL1252
• Backbone manufacturer: Naweed Lab
• Backbone size w/o insert (bp): 6845
• Total vector size (bp): 10687
• Vector type: Fungal expression (in Magnaporthe oryzae)
• Selectable markers: Hygromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: tTA+Tnos
• Species: Synthetic
• Insert Size (bp): 1182
• Mutation: Codon optimized for Magnaporthe oryzae
• Promoter: promoter less

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoR I (not destroyed)
• 3′ cloning site: Sal I (not destroyed)
• 5′ sequencing primer: CGCGAATTCATGGCCCGCCTGGATAAGTCAAAG
• 3′ sequencing primer: CAGGTCGACGCGGCCGCAATTCTCATGTTTGACA

Gene/Insert 2

• Gene/Insert name: Hygromycin cassette
• Alt name: hptII
• Species: E. coli
• Insert Size (bp): 2008
• Promoter: Aspergillus nidulans TrpC promoter

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xba I (not destroyed)
• 3′ cloning site: Sal I (not destroyed)
• 5′ sequencing primer: CCGTCTAGAGTCGAGGATATTGAAGGAGCACT
• 3′ sequencing primer: CGCGTCGACAAGAAGGATTACCTCTAAAC

Gene/Insert 3

• Gene/Insert name: Ttub+PtetO
• Species: Synthetic
• Insert Size (bp): 682
• Promoter: Ustilago maydis Mfa1 basal promoter with six tetO binding sites

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xba I (not destroyed)
• 3′ cloning site: Kpn I (not destroyed)
• 5′ sequencing primer: GACTCTAGATAAATCATTCCACTCAACATTCAG
• 3′ sequencing primer: CCCGGTACCCATATTAATTGATAGAAGT

Terms and Licenses

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