pKCcas9dO
(Plasmid #62552)

• Purpose: High efficiency Streptomyces genome editing by CRISPR/Cas9 system
• Depositing Lab: Weihong Jiang
• Publication: Huang et al Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi: 10.1093/abbs/gmv007. Epub 2015 Mar 3.

Backbone

• Vector backbone: pKC1139
• Vector type: Bacterial Expression, CRISPR ; Bacterial Genome Editing

Growth in Bacteria

• Bacterial Resistance(s): Apramycin, 25 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: Scocas9
• Species: artificially synthesized, codon optimized gene from Strep pyogenes
• Insert Size (bp): 4107
• Promoter: ptipA

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: EcoRI (unknown if destroyed)
• 5′ sequencing primer: unknown
• 3′ sequencing primer: M13pUC-Rev

Gene/Insert 2

• Gene/Insert name: sgRNA ()
• Insert Size (bp): 103
• Promoter: j23119

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: j23119-Fwd

Gene/Insert 3

• Gene/Insert name: act-orf4 homology arm
• Insert Size (bp): 2013

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: unknown
• 3′ sequencing primer: M13pUC-Fwd

Resource Information

• Supplemental Documents:
  • Genbank
  • Supplemental Information
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

More information is provided in the supplemental files. Please note that the sequence of the backbone (pKC1139) was obtained via Sanger sequencing and several regions may have discrepancies due to high GC content. These discrepancies have no functional consequences.