pAH-CTX1-rhadCas9
(Plasmid #129391)

• Purpose: Derived from pAH-CTX1-rha. The Burkholderia cenocepacia codon-optimized dCas9 cloned downstream of PrhaBAD. Hence, dCas9 is controlled by the rhamnose-inducible promoter system.
• Depositing Lab: Silvia Cardona
• Publication: Hogan et al ACS Synth Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232.

Backbone

• Vector backbone: pAH-CTX1-rha
• Backbone manufacturer: Andrew Hogan
• Backbone size w/o insert (bp): 7636
• Total vector size (bp): 11897
• Modifications to backbone: Insertion of codon-optimized dCas9 for Burkholderia cenocepacia.
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Tetracycline, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Burkholderia cenocepacia codon-optimized dCas9
• Species: Synthetic; S. pyogenes
• Insert Size (bp): 4247
• Mutation: Entire dCas9 codon optimized for the GC-rich B. cenocepacia. See this sequence and that of pAH-CTX1-rhadCas9-native for comparison.
• GenBank ID: AE004092.2
• Promoter: PrhaBAD

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: ATTTTTGCGGCGCGTGTAG
• 3′ sequencing primer: GGGTTCTATCGCCACGGAC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Codon-optimized dCas9 was synthesized in two parts by IDT. Using ligation and PCR amplification, the full dCas9 vector was constructed and subsequently cloned into the pAH-CTX1-rha background. This insert was designed based on the dCas9 CDS from pdCas9-bacteria (Qi et al. 2013).

Terms and Licenses

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