pPV033
(Plasmid #178359)

• Purpose: Targeted gene knock-in plasmid for Schizophyllum commune
• Depositing Lab: Han Wösten
• Publication: Vonk et al Open Res Europe 2021, 1:140

Backbone

• Vector backbone: pRO402
• Backbone manufacturer: Robin Ohm
• Backbone size w/o insert (bp): 4173
• Total vector size (bp): 7567
• Modifications to backbone: Integration of two homology arms and a nourseothricin resistance cassette
• Vector type: Fungal homologous recombination plasmid
• Selectable markers: Zeocin ; Nourseothricin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Left flank
• Species: Schizophyllum commune
• Insert Size (bp): 1060
• Promoter: n.a.

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ctatgaccatgattacgccaGCCAACAATCAGATCCGCTAGC
• 3′ sequencing primer: tcccagaccaccatgccgggCCATGGAGCAACCTCGGTGCTTATGACT

Gene/Insert 2

• Gene/Insert name: nour_cassette
• Alt name: Nourseothricin resistance cassette
• Species: Synthetic; Schizophyllum commune
• Insert Size (bp): 1307
• Promoter: S. commune gpd

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n.a.
• 3′ sequencing primer: n.a.

Gene/Insert 3

• Gene/Insert name: Right flank
• Species: Schizophyllum commune
• Insert Size (bp): 1067
• Promoter: n.a.

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: gtccccctcgaggcgcgccgGGGTCCTTTCTGACGATGACCA
• 3′ sequencing primer: gataaccttcagcagaacgaCGTGACTTTTCGCCGTCTGTAC

Resource Information

• Supplemental Documents:
  • ppv033-targeted-insertion.gb

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.12688/openreseurope.14262.1 for preprint available on Open Research Europe.