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David Sibley Lab Plasmids


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Title Authors Publication
Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9. Shen B, Brown KM, Lee TD, Sibley LD MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14.
The Toxoplasma pseudokinase ROP5 forms complexes with ROP18 and ROP17 kinases that synergize to control acute virulence in mice. Etheridge RD, Alaganan A, Tang K, Lou HJ, Turk BE, Sibley LD Cell Host Microbe. 2014 May 14;15(5):537-50. doi: 10.1016/j.chom.2014.04.002.
Genetic mapping reveals that sinefungin resistance in Toxoplasma gondii is controlled by a putative amino acid transporter locus that can be used as a negative selectable marker. Behnke MS, Khan A, Sibley LD Eukaryot Cell. 2014 Dec 5. pii: EC.00229-14.
Analysis of non-canonical calcium dependent protein kinases in Toxoplasma gondii by targeted gene deletion using CRISPR/Cas9. Long S, Wang Q, Sibley LD Infect Immun. 2016 Jan 11. doi: 10.1128/IAI.01173-15
pLinker-6xHA-HXGPRT-LoxP (unpublished)
Plasma Membrane Association by N-Acylation Governs PKG Function in Toxoplasma gondii. Brown KM, Long S, Sibley LD MBio. 2017 May 2;8(3). pii: e00375-17. doi: 10.1128/mBio.00375-17.
Calmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii. Long S, Brown KM, Drewry LL, Anthony B, Phan IQH, Sibley LD PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379.
ID Plasmid Gene/Insert Vector Type Publication Hidden Extra Search Info  
54467pSAG1::CAS9-U6::sgUPRTSAG1 5' UTR (Other)CRISPR ; Toxoplasma expressionEfficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9. MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14. Encodes CAS9 nuclease (GFP fusion) under control of the Toxoplasma gondii SAG1 promotor. Vector also provides U6 controlled expression of a single guide RNA for disruption of genes by CRISPR. pBlueScript Add to Cart
58528pUPRT::DHFR-DDHFR cassette (Other)Gateway cloningEfficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9. MBio. 2014 May 13;5(3). pii: e01114-14. doi: 10.1128/mBio.01114-14. replace TgUPRT with DHFR by homologous recombination pDEST R4-R3 Vector II Add to Cart
59017pTet-On-TAPToxoplasma ExpressionThe Toxoplasma pseudokinase ROP5 forms complexes with ROP18 and ROP17 kinases that synergize to control acute virulence in mice. Cell Host Microbe. 2014 May 14;15(5):537-50. doi: 10.1016/j.chom.2014.04.002. Construct allows tetracycline inducible expression of a protein of interest fused to a tandem affinity purification tag (TAP) in toxoplasma gondii. pTet07Sag1mycGFP
  • Tag / Fusion Protein
    • Calmodulin Binding Peptide fused with a TEV Cleavage site to Protein A-tag (C terminal on backbone)
  • Add to Cart
    59018pCAT-YFP-TetRYFP-TetR (Other)Toxoplasma ExpressionThe Toxoplasma pseudokinase ROP5 forms complexes with ROP18 and ROP17 kinases that synergize to control acute virulence in mice. Cell Host Microbe. 2014 May 14;15(5):537-50. doi: 10.1016/j.chom.2014.04.002. Expresses a fusion of yellow fluorescent protein (YFP) to the N-terminus of the Escherichia coli Tn10 tet repressor (TetR) driven by the alpha tubulin promoter for use in T. gondii pCAT-GFP Add to Cart
    59855pSAG1::CAS9-U6::sg290860-6CRISPR sg290860-6CRISPR ; ; Toxoplasma gondiiGenetic mapping reveals that sinefungin resistance in Toxoplasma gondii is controlled by a putative amino acid transporter locus that can be used as a negative selectable marker. Eukaryot Cell. 2014 Dec 5. pii: EC.00229-14. Encodes CAS9 nuclease (GFP fusion) under control of the Toxoplasma gondii SAG1 promotor. Vector also provides U6 controlled expression of a single guide RNA for CRISPR disruption of TGME49_290860. pBlueScript Add to Cart
    70147pLoxP-DHFR-mCherrya selectable dihydrofolate reductase-thymidylate synthase marker (Other)Expression in Toxoplasma gondiiAnalysis of non-canonical calcium dependent protein kinases in Toxoplasma gondii by targeted gene deletion using CRISPR/Cas9. Infect Immun. 2016 Jan 11. doi: 10.1128/IAI.01173-15 Expresses a DHFR-mCherry fusion protein and uses for a CRISPR gene deletion pUC19 Add to Cart
    86552pLinker-6xHA-HXGPRT-LoxPLinker-6xHA-HGPRT-LoxP (Other)Bacterial Expression pLinker-6xHA-HXGPRT-LoxP (unpublished) The plasmid is used for endogenous tagging using CRISPR technology in Toxoplasma pUC19 Add to Cart
    86553pLinker-AID-3xHA-HXGPRT-LoxPLinker-AID-3xHA-HXGPRT 3'UTR (Other)Bacterial ExpressionCalmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii. PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379. used for a TIR1-AID system with CRISPR tagging technology in Toxoplasma gondii pUC19 Add to Cart
    86667pLinker-AID-Ty-HXGPRT-LoxPLinker-AID-Ty-HGPRT 3'UTR (Other)Bacterial ExpressionCalmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii. PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379. for generation of AID strains using CRISPR tagging technology pUC19 Add to Cart
    86669pLinker-AID-3xHA-DHFR-LoxPLinker-AID-3xHA-HXGPRT 3'UTR (Other)Bacterial ExpressionCalmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii. PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379. for generation of AID strains using CRISPR tagging technology pUC19 Add to Cart
    87258pTUB1:OsTIR1-3FLAG, SAG1:CATTIR1 (Synthetic), CAT (Synthetic)Toxoplasma expressionPlasma Membrane Association by N-Acylation Governs PKG Function in Toxoplasma gondii. MBio. 2017 May 2;8(3). pii: e00375-17. doi: 10.1128/mBio.00375-17. TIR1-3FLAG fusion driven by a TUB1 promoter with a CAT drug selectable marker. The TIR1 CDS from Oryza sativa was codon optimized for T. gondii expression. pBluescript SK(+) Add to Cart
    87259pTUB1:YFP-mAID-3HA, DHFR-TS:HXGPRTYFP-mAID-3HA (Synthetic), HXGPRT (Other)Toxoplasma expressionPlasma Membrane Association by N-Acylation Governs PKG Function in Toxoplasma gondii. MBio. 2017 May 2;8(3). pii: e00375-17. doi: 10.1128/mBio.00375-17. YFP-mAID-3HA fusion driven by a minimal TUB1 promoter with an HXGdrug selectable marker. The mAID CDS is from Arabidopsis thaliana auxin-responsive protein IAA17E66-S133 pBluescript SK(+) Add to Cart
    87260pTUB1:YFP-AID-3HA, DHFR-TS:HXGPRTYFP-AID-3HA (Synthetic), HXGPRT (Other)Toxoplasma expressionPlasma Membrane Association by N-Acylation Governs PKG Function in Toxoplasma gondii. MBio. 2017 May 2;8(3). pii: e00375-17. doi: 10.1128/mBio.00375-17. YFP-AID-3HA fusion driven by a minimal TUB1 promoter with an HXGPRT drug selectable marker. The AID CDS is Arabidopsis thaliana auxin-responsive protein IAA17. pBluescript SK(+) Add to Cart