pCas9-CAT
(Plasmid
#80323)
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PurposeEncodes Cas9 and chloramphenicol acetyltransferase (CAT)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 80323 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepU6-Universal
- Backbone size w/o insert (bp) 8360
- Total vector size (bp) 9301
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Modifications to backboneThe sgRNA region has been replaced with chloramphenicol acetyltransferase with the TUB1 5'UTR and the SAG1 3' UTR
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Vector typeCRISPR
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Selectable markersCAT
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert 1
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Gene/Insert nameCas9
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SpeciesStreptococcus pyogenes
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Insert Size (bp)4101
- Promoter TgTUB1
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Tags
/ Fusion Proteins
- 3X FLAG (N terminal on insert)
- NLS (N terminal on insert)
- NLS (C terminal on insert)
Cloning Information for Gene/Insert 1
- Cloning method Unknown
- 5′ sequencing primer CAT-F
- 3′ sequencing primer BGH-rev (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameChloramphenicol acetyltransferase
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Alt nameCAT
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Insert Size (bp)1535
- Promoter TgTUB1
Cloning Information for Gene/Insert 2
- Cloning method Restriction Enzyme
- 5′ cloning site PciI (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer pBluescript-KS
- 3′ sequencing primer pBluescript-SK (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
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.
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:
pCas9-CAT was a gift from Sebastian Lourido (Addgene plasmid # 80323 ; http://n2t.net/addgene:80323 ; RRID:Addgene_80323) -
For your References section:
A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, Saeij JP, Carruthers VB, Niles JC, Lourido S. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. 10.1016/j.cell.2016.08.019 PubMed 27594426