pBFC1207
(Plasmid #186694)

• Purpose: VcDART under control of Lac promoter, Vc_2xBsaI_NT gRNA, GmR+sfGFP cargo, with ET-Seq (AsiSI+SbfI) restriction sites flanking Tn7 Right End
• 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: tnsA
• Species: Vibrio cholerae
• Promoter: PLac

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: tnsB
• Species: Vibrio cholerae

Gene/Insert 3

• Gene/Insert name: tnsC
• Species: Vibrio cholerae

Gene/Insert 4

• Gene/Insert name: tnsD
• Species: Vibrio cholerae

Gene/Insert 5

• Gene/Insert name: tniQ
• Species: Vibrio cholerae

Gene/Insert 6

• Gene/Insert name: cas8-cas5 fusion
• Species: Vibrio cholerae

Gene/Insert 7

• Gene/Insert name: cas7
• Species: Vibrio cholerae

Gene/Insert 8

• Gene/Insert name: cas6
• Species: Vibrio cholerae

Gene/Insert 9

• Gene/Insert name: Vc_2xBsaI_NT (Guide Stuffer) crRNA
• Species: Vibrio cholerae

Gene/Insert 10

• Gene/Insert name: Tn7 transposon
• Species: Vibrio cholerae

Gene/Insert 11

• Gene/Insert name: GmR+sfGFP VcDART Cargo
• Alt name: gentamicin marker+sfGFP VcDART Cargo

Gene/Insert 12

• Gene/Insert name: SbfI+AsiSI dual restriction site for donor plasmid removal during ET-Seq

Resource Information

• Supplemental Documents:
  • pBFC1207 clean for addgene.gbk

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.
This vector is identical to pBFC0619, but it includes a dual restriction site (AsiSI+SbfI) flanking the Tn right end to reduce donor vector during ET-Seq.