pEC2483 (pSpy1C)
(Plasmid
#220248)
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Purpose(Empty Backbone) replicative E. coli-S. pyogenes shuttle plasmid for heterologous gene expression
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 220248 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepSpy1C
- Backbone size (bp) 4917
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Modifications to backboneSee associated article for detailed construction information.
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsFor E. coli, use 20 µg/mL chloramphenicol in LB medium. For S. pyogenes, use 20 µg/mL chloramphenicol in THY medium.
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Copy numberHigh Copy
Cloning Information
- Cloning method Golden Gate
- 5′ sequencing primer GTAAAACGACGGCCAGTC
- 3′ sequencing primer CAGGAAACAGCTATGAC (Common Sequencing Primers)
Resource Information
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Supplemental Documents
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
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.
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.
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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