pBFC0687
(Plasmid #186692)

• Purpose: ShDART under control of Lac promoter, original configuration (pre-gRNA terminator and J23119 promoter), Sh_2xBsaI_NT gRNA, GmR+sfGFP cargo
• Depositing Lab: Jennifer Doudna
• Publication: Rubin et al Nat Microbiol. 2022 Jan;7(1):34-47. doi: 10.1038/s41564-021-01014-7. Epub 2021 Dec 6.

Backbone

• Vector backbone: R6K conditional origin, bla, RP4 oriT
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Gentamicin, 100 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): EC100D pir+
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: tnsB
• Species: Scytonema hofmanni
• Promoter: PLac

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Gene/Insert 2

• Gene/Insert name: tnsC
• Species: Scytonema hofmanni

Gene/Insert 3

• Gene/Insert name: tniQ
• Species: Scytonema hofmanni

Gene/Insert 4

• Gene/Insert name: Cas12k
• Alt name: ShCas12k
• Species: Scytonema hofmanni

Gene/Insert 5

• Gene/Insert name: Sh_2xBsaI_NT (Guide Stuffer) gRNA
• Species: Scytonema hofmanni

Gene/Insert 6

• Gene/Insert name: Tn7 transposon
• Species: Scytonema hofmanni

Gene/Insert 7

• Gene/Insert name: GmR+sfGFP ShDART Cargo
• Alt name: gentmycin marker+sfGFP ShDART Cargo

Resource Information

• Supplemental Documents:
  • pBFC0687 clean for addgene.gbk
• A portion of this plasmid was derived from a plasmid made by: Feng Zhang lab's ShCAST plasmids: Addgene #127921 and #127924

Terms and Licenses

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

Depositor Comments

Clone guides using the 2xBsaI cloning site.
Clone Tn barcodes using the 2xBsmBI cloning site.