pCRI003-pGEM-LS-Bar-PgpdA-dSpCas9-VPR-TtrpC-LS
(Plasmid #140196)

• Purpose: Chromosomal integration of PgpdA-dSpCas9-VPR-TtrpC. Contains 1kb homology arms for Aspergillus nidulans and the bar gene for glufosinate selection. Filamentous fungi vector.
• Depositing Lab: Yit Heng Chooi
• Depositing Organization: University of Western Australia
  Located in Australia
• Publication: Roux et al ACS Synth Biol. 2020 Jun 11. doi: 10.1021/acssynbio.0c00197.

Backbone

• Vector backbone: pGem
• Backbone size w/o insert (bp): 3033
• Total vector size (bp): 12892
• Vector type: CRISPR, Synthetic Biology ; Fungal genome integration.
• Selectable markers: Basta

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: dSpCas9-VPR
• Alt name: dCas9-VPR
• Species: Synthetic; Streptococcus pyogenes
• Insert Size (bp): 5730
• Promoter: PgpdA
• Tag / Fusion Protein:
  • VPR (VP64-p65-Rta) , NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GTCAGTCCAACATTTGTTGC

Gene/Insert 2

• Gene/Insert name: Bar
• Alt name: Glufosinate/basta-resistance
• Species: Streptomyces hygroscopicus
• Insert Size (bp): 552
• Promoter: PtrpC

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GTCGACAGAAGATGATATTGAAGGAG

Gene/Insert 3

• Gene/Insert name: Homology arm (ChrIV_A_nidulans_FGSC_A4:2736011,2737053)
• Species: Aspergillus nidulans
• Insert Size (bp): 1040

Gene/Insert 4

• Gene/Insert name: Homology arm (ChrIV_A_nidulans_FGSC_A4:2790295,2791327)
• Species: Aspergillus nidulans
• Insert Size (bp): 1033

Terms and Licenses

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

Depositor Comments

To be linerised with NotI for homologous recombination in Aspergillus nidulans nkuA deficient strains, selecting with glufosinate/Basta. It can also be used for random genome integration in other fungal strains.

DNA (Catalog # 140196-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 140196-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.