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PurposeExpresses the fusion gene CAS9-HA-2xNLS-GFP in the pTREX-n backbone. This vector is used for cloning a specific sgRNA by BamHI, to be co-expressed with Cas9 for genome editing in Trypanosoma cruzi.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 68708 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepTREX-n
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Backbone manufacturerVazquez and Levin., 1999
- Backbone size w/o insert (bp) 6227
- Total vector size (bp) 11173
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Modifications to backboneCloning of gene Cas9-HA-2xNLS-GFP mutated (C4239T) into pTREX-n vector
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Vector typeCRISPR
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Selectable markersNeomycin (select with G418)
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 numberHigh Copy
Gene/Insert
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Gene/Insert nameFusion gene Cas9-HA-2xNLS-GFP from vector pMJ920 (Addgene) .
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Alt nameCas9-GFP
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SpeciesSynthetic; Streptococcus pyogenes
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Insert Size (bp)4975
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MutationC4239T mutation that eliminates a BamHI restriction site on the Cas9-GFP ORF.
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Tag
/ Fusion Protein
- Fusion gene Cas9-HA-2xNLS-GFP from vector pMJ920 (Addgene)
Resource Information
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A portion of this plasmid was derived from a plasmid made byThe gene was cloned by Dr. Noelia Lander in my laboratory. The sequence was obtained from vector pMJ920 (Addgene), kindly provided by Dr. Vasant Muralidharan (University of Georgia, Athens, GA).
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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:
Cas9/pTREX-n was a gift from Roberto Docampo (Addgene plasmid # 68708 ; http://n2t.net/addgene:68708 ; RRID:Addgene_68708) -
For your References section:
CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 Genes in Trypanosoma cruzi Reveals Their Role in Flagellar Attachment. Lander N, Li ZH, Niyogi S, Docampo R. MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15. 10.1128/mBio.01012-15 PubMed 26199333