pL2Cas9
(Plasmid #98841)

• Purpose: Genome engineering of virulent phages
• Depositing Lab: Sylvain Moineau
• Publication: Lemay et al ACS Synth Biol. 2017 Mar 30. doi: 10.1021/acssynbio.6b00388.

Backbone

• Vector backbone: pTRKL2
• Backbone size w/o insert (bp): 6419
• Total vector size (bp): 12221

Growth in Bacteria

• Bacterial Resistance(s): Erythromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB5-alpha
• Growth instructions: Growth medium: BHI + Erythromycin 150 µg/ml.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: tracr/Cas9
• Insert Size (bp): 5802

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GCATATTAAACTAATTTCGGAGGTC
• 3′ sequencing primer: CTGTATTACTGCATTTATTAAGAGTATTATACC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The original gene was cloned by Luciano Marrafini and received from Addgene #42876.

Terms and Licenses

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

Depositor Comments

Note from the paper of O'Sullivan, and Klaenhammer: We found that BHI (Difco Laboratories, Detroit, MI, USA) was an excellent selection medium for ErR in E. coli. Unlike LB medium, BHI plates containing 150µg Er/ml afforded a clean selection, with no background colonies appearing even upon prolonged incubaction of one week or more.

O'Sullivan, D.J., T.R. Klaenhammer. 1993. High- and low-copy-number Lactococcus shuttle cloning vectors with features for clone screening. Gene 137:227-231.