pPfetFn-Target_v2
(Plasmid #199564)

• Purpose: gRNA expression for FnCas12a in E. coli and clostridia
• Depositing Lab: Philippe Lambin
• Publication: Zhang et al Microbiol Spectr. 2023 Nov 10:e0245923. doi: 10.1128/spectrum.02459-23.

Backbone

• Vector backbone: pGG2121
• Backbone size w/o insert (bp): 1500
• Total vector size (bp): 6365
• Modifications to backbone: Based on pMTL82121, change teminator TCD0164 to TtyrS and add BsaI Golden-Gate sites
• Vector type: Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: SacB
• gRNA/shRNA sequence: AATTTCTACTGTTGTAGATtGAGACGattCGTCTCcAATTTCTACTGTTGTAGAT
• Species: Bacillus subtilis
• GenBank ID: 936413
• Promoter: Pthl_fU

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PvuI (not destroyed)
• 3′ cloning site: PstI (not destroyed)
• 5′ sequencing primer: GCcgatcgTTTTTAACAAAATATATTGATAAAAATAATAATAGTGGGT
• 3′ sequencing primer: cgctgcagttatttgttaactgttaattgtccttg

Resource Information

• Supplemental Documents:
  • copy-of-ppfetfn-target_v2.gb

Terms and Licenses

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