pPEE17
(Plasmid #128361)

• Purpose: Detecting gene essentiality in Cryptococcus neoformans. Targets DNA constructs to the Safe Haven Site 1, a small gene-free region. Contains amdS2 counterselectable marker. This version of the Safe Haven vector includes direct repeats, enabling the formation of popouts on fluoroacetamide.
• Depositing Lab: James Fraser
• Depositing Organization: University of Queensland
  Located in Australia
• Publication: Erpf et al Fungal Genet Biol. 2019 Jun 17;131:103241. doi: 10.1016/j.fgb.2019.103241.

Backbone

• Vector backbone: pBlueScript II SK(-)
• Backbone manufacturer: Stratagene
• Vector type: Unspecified
• Selectable markers: Acetamide, Fluoroacetamide

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: Acetamidase (amdS)
• Species: Synthetic
• Promoter: TEF1

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

Contains the Safe Haven Site 1 sequence (CNAG_00777 - CNAG_00778) in the backbone of pBluescript II SK- flanked by blaster repeats to enable integration into the C. neoformans genome via homologous recombination, in addition to a linker sequence containing multiple rare-cutting restriction enzyme cut sites to facilitate linearization prior to biolistic transformation. Importantly, as the Safe Haven and marker fragments were inserted into the pBluescript II SK- backbone in place of the f1 origin of replication, each plasmid still retains the original multicloning site and blue/white screening capabilities. Linearization of the safe haven constructs is made possible by the inclusion of a linker containing cut sites for three rare cutting restriction enzymes, BaeI, AscI and PacI. To use the vectors, the gene of interest is subcloned into the standard pBluescript II SK- multicloning site.

DNA (Catalog # 128361-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 # 128361-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.