pBWB424
(Plasmid #140638)

• Purpose: RSF1010-Cas9-vanAB gRNA. Vector with constitutive Cas9 and gRNA targeting vanAB locus.
• Depositing Lab: Keith Tyo
• Publication: Biggs et al Nucleic Acids Res. 2020 Apr 4. pii: 5815822. doi: 10.1093/nar/gkaa167.

Backbone

• Vector backbone: CRISPRi-ME (Jeffrey Barrick Laboratory)
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Can be grown at 30 or 37 C. 30 C used for referenced study in ADP1, del_vanAB::kanR-lacI-Trc-mCherry.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Cas9
• Insert Size (bp): 4107

Resource Information

• Supplemental Documents:
  • rsf1010-t5-sgrna-vanab-1-tdk-cas9 (1).gb
• A portion of this plasmid was derived from a plasmid made by: Received from Jeffrey Barrick laboratory at UT Austin.

Terms and Licenses

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

Depositor Comments

Derivative of CRISPRi-ME from Peng et al., "Synthetic Genome Defenses against Selfish DNA Elements Stabilize Engineered Bacteria against Evolutionary Failure", ACS Synthetic Biology, 2019. Vector was modified to take the dCas9 to Cas9 and gRNA was substituted to no longer target the insertion sequences, but instead the vanAB genes in ADP1.