pFD115
(Plasmid #124768)

• Purpose: pFD115 carries cas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector
• Depositing Lab: David Bikard
• Publication: Depardieu et al Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024.

Backbone

• Vector backbone: pLZ12
• Backbone size w/o insert (bp): 3729
• Total vector size (bp): 9131
• Vector type: CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: Ptet
• Insert Size (bp): 855
• Promoter: Ptet

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AAGCACCCATTAGTTCAGTTACCATCACGGAAAAAGGTTATGCTG
• 3′ sequencing primer: CCATTTTTGCCTCCTATATGCCTCCGAATTCGATATCACT

Gene/Insert 2

• Gene/Insert name: cas9
• Species: Streptococcus pyogenes

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GAATTCGGAGGCATATAGGAGGCAAAAATGGATAAGAAATA
• 3′ sequencing primer: TCAGTCACCTCCTAGCTGACTCAA

Gene/Insert 3

• Gene/Insert name: term PpflB sgRNA (BsaI)
• Species: Synthetic

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GTCTTTATGAAACACGCATTGA
• 3′ sequencing primer: ATCCAATTTTCGTTTGTAGTAGAAACAGACGAAGAATCCATG

Resource Information

• Supplemental Documents:
  • pFD115.gb

Terms and Licenses

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