BB3cN_pGAP_23*_pPFK300_Cas9
(Plasmid
#104911)
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PurposehCas9 under control of pPFK300 for direct cloning of HH-sgRNA-HDV PCR products for episomal expression in P. pastoris and selection on NTC
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 104911 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneEpisomal ARS/CEN Backbone
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Vector typeYeast Expression, CRISPR
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Selectable markersNTC
Growth in Bacteria
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Bacterial Resistance(s)Nourseothricin (clonNat), 50 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 1
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Gene/Insert nameHH - FS23 linker - HDV
Cloning Information for Gene/Insert 1
- Cloning method Restriction Enzyme
- 5′ cloning site BbsI (unknown if destroyed)
- 3′ cloning site BbsI (not destroyed)
- 5′ sequencing primer pGAP fw (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert namehuman codon-optimized hcas9 sequence fused to a nuclear localization signal (NLS)
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Alt namehcas9
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SpeciesH. sapiens (human); Streptococcus pyogenes, Pichia pastoris
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Insert Size (bp)4140
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byThe original Cas9 sequence was taken from from DiCarlo et al., (Addgene ID 43802). Reference: DiCarlo JE, Norville JE, Mali P, Rios X, Aach J, Church GM. 2013. Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res 41:4336-43.
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
Additional Publication: Prielhofer, R. etal (2017) BMC Syst Biol. 11,123 (PMID:29221460)
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:
BB3cN_pGAP_23*_pPFK300_Cas9 was a gift from Brigitte Gasser (Addgene plasmid # 104911 ; http://n2t.net/addgene:104911 ; RRID:Addgene_104911) -
For your References section:
CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia pastoris. Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R. Methods Mol Biol. 2019;1923:211-225. doi: 10.1007/978-1-4939-9024-5_9. 10.1007/978-1-4939-9024-5_9 PubMed 30737742