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Addgene

pEC2483 (pSpy1C)
(Plasmid #220248)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 220248 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pSpy1C
  • Backbone size (bp) 4917
  • Modifications to backbone
    See associated article for detailed construction information.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    For E. coli, use 20 µg/mL chloramphenicol in LB medium. For S. pyogenes, use 20 µg/mL chloramphenicol in THY medium.
  • Copy number
    High Copy

Cloning Information

  • Cloning method Golden Gate
  • 5′ sequencing primer GTAAAACGACGGCCAGTC
  • 3′ sequencing primer CAGGAAACAGCTATGAC
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The multiple cloning site (EcoRI, XbaI, SpeI and PstI) contains the Plac_mrfp cassette. The mrfp reporter allows for red-white screening during sub-cloning in E. coli (DH5a). When the Plac_mrfp cassette is replaced with the insert of interest, colonies will appear white.

Resource Information: We took the rolling circle replicon pSH71 and the chloramphenicol resistance cassette from pIB166 (Plasmid #90189). The the Plac_mrfp cassette for the MCS is derived from pBS3Clux (Plasmid #55172).

Please download the file linked below to obtain a well annotated map of the plasmid.

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:

    pEC2483 (pSpy1C) was a gift from Emmanuelle Charpentier (Addgene plasmid # 220248 ; http://n2t.net/addgene:220248 ; RRID:Addgene_220248)
  • For your References section:

    Expanding the genetic toolbox for the obligate human pathogen Streptococcus pyogenes. Lautenschlager N, Schmidt K, Schiffer C, Wulff TF, Hahnke K, Finstermeier K, Mansour M, Elsholz AKW, Charpentier E. Front Bioeng Biotechnol. 2024 Jun 7;12:1395659. doi: 10.3389/fbioe.2024.1395659. eCollection 2024. 10.3389/fbioe.2024.1395659 PubMed 38911550