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Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens.
Mallett DR, Chang M, Cheng X, Bezanilla M
Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep.
PubMed Article

These materials are included in the CRISPR-Cas9 Gateway System for Physcomitrella patens Kit.

Plasmids from Article

ID Plasmid Purpose  
113735pENTR-PpU6P-sgRNA-L1L2Gateway entry vector containing attL1 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113736pENTR-PpU6P-sgRNA-L1L4Gateway entry vector containing attL1 and attL4 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113737pENTR-PpU6P-sgRNA-L1R5Gateway entry vector containing attL1 and attR5 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113738pENTR-PpU6P-sgRNA-L5L2Gateway entry vector containing attL5 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113739pENTR-PpU6P-sgRNA-R4R3Gateway entry vector containing attR4 and attR3 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113740pENTR-PpU6P-sgRNA-L3L2Gateway entry vector containing attL3 and attL2 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113741pENTR-PpU6P-sgRNA-L5L4Gateway entry vector containing attL5 and attL4 sites, U6 promoter, BsaI sites for protospacer ligation, and sgRNA.
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113742pMH-Cas9-gateGateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving hygromycin for selection in plants.
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113743pMK-Cas9-gateGateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving aminoglycoside phosphotransferase for G418 selection in plants.
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113744pZeo-Cas9-gateGateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving Zeocin for selection in plants.
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113745pENTR-R4R3-stopContains three stop codons in every frame to ensure a premature stop codon when introduced into the genome via homology-directed repair. Gateway multisite entry clone.
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113746pENTR-R4R3-mEGFP-NGateway multi-site entry clone for second position mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113747pENTR-R4R3-2xmEGFP-NGateway multi-site entry clone for second position 2x mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113748pENTR-R4R3-3xmEGFP-NGateway multi-site entry clone for second position 3x mEGFP (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113749pENTR-R4R3-mEGFP-CGateway multi-site entry clone for second position mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113750pENTR-R4R3-2xmEGFP-CGateway multi-site entry clone for second position 2x mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113751pENTR-R4R3-3xmEGFP-CGateway multi-site entry clone for second position 3x mEGFP (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113752pENTR-R4R3-mRuby-NGateway multi-site entry clone for second position mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113753pENTR-R4R3-2xmRuby-NGateway multi-site entry clone for second position 2x mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113754pENTR-R4R3-3xmRuby-NGateway multi-site entry clone for second position 3x mRuby (N-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113755pENTR-R4R3-mRuby-CGateway multi-site entry clone for second position mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113756pENTR-R4R3-2xmRuby-CGateway multi-site entry clone for second position 2x mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113757pENTR-R4R3-3xmRuby-CGateway multi-site entry clone for second position 3x mRuby (C-terminal tag). Introduced into the genome via homology-directed repair after an LR reaction with homology sequences.
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113758pGEM-gateGateway destination vector only containing the Gateway cassette and pGEM-T Easy backbone. This can be used with Gateway-compatible Homology-Directed Repair.
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Antibodies from Article