pCas9GG
(Plasmid #131009)

• Purpose: Backbone plasmid for generating CRISPR arrays for SpCas9 using CRATES. Contains a direct repeat and a RFP-dropout cassette.
• Depositing Lab: Chase Beisel
• Publication: Liao et al Nat Commun. 2019 Jul 3;10(1):2948. doi: 10.1038/s41467-019-10747-3.

Backbone

• Vector backbone: pBAD18
• Total vector size (bp): 4287
• Modifications to backbone: arabinose PBAD promoter is replaced by PJ23119 promoter
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: direct repeat of SpCas9
• Insert Size (bp): 36

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AGCGCTCATGAGCCCGA
• 3′ sequencing primer: TGCCGCCAGGCAAATTCTG

Gene/Insert 2

• Gene/Insert name: promoter PJ23119
• Insert Size (bp): 35

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AGCGCTCATGAGCCCGA
• 3′ sequencing primer: TGCCGCCAGGCAAATTCTG

Gene/Insert 3

• Gene/Insert name: mRFP expression cassette
• Insert Size (bp): 895
• Promoter: Bba_R0040 TetR

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AGCGCTCATGAGCCCGA
• 3′ sequencing primer: TGCCGCCAGGCAAATTCTG

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry