Skip to main content
This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected]. Learn more

pPEE21
(Plasmid #128364)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 128364 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

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

Resource Information

  • Supplemental Documents

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Contains the Safe Haven Site 5 sequence (CNAG_05670 - CNAG_05671) in the backbone of pBluescript II SK- 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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pPEE21 was a gift from James Fraser (Addgene plasmid # 128364 ; http://n2t.net/addgene:128364 ; RRID:Addgene_128364)
  • For your References section:

    amdS as a dominant recyclable marker in Cryptococcus neoformans. Erpf PE, Stephenson CJ, Fraser JA. Fungal Genet Biol. 2019 Jun 17;131:103241. doi: 10.1016/j.fgb.2019.103241. 10.1016/j.fgb.2019.103241 PubMed 31220607