pIA030
(Plasmid #200424)

• Purpose: thrC::(Ppen-lacIΔ11-PAmCherry1 erm2). PALM integration vector includes PAmCherry1 fusion tagged C-terminal of LacI for FROS imaging in B. subtilis.
• Depositing Lab: Rut Carballido-López
• Publication: Altinoglu et al bioRxiv 2023

Backbone

• Vector backbone: pIA028
• Backbone size w/o insert (bp): 6806
• Total vector size (bp): 8099
• Vector type: Bacterial Expression
• Selectable markers: Erythromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: E. coli: 37°C and selection ampicillin 100 μg/mL B. subtilis: 30°C and selection erythromycin 0.5 μg/mL
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: LacI
• Species: Escherichia coli
• Insert Size (bp): 1047
• GenBank ID: J01636.1
• Promoter: Ppen
• Tag / Fusion Protein:
  • PAmCherry (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CGAATTCGAGCTCGGTACGGGGATCGGGTTATCGAATTCCCGGTGG
• 3′ sequencing primer: CCTCCTCGCCCTTGCTCACGGCGGCCGCCAGCTGCATTAATGAATCGGCC

Resource Information

• Supplemental Documents:
  • pIA025 aka ThrC-LacI-bsPamCherry.gbk

Terms and Licenses

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

Depositor Comments

pPT300 was amplified using primers oIA027 and oIA028. The resulting fragment was cloned into pIA028 digested with BamHI and NotI, giving pIA030, thrC::(Ppen-lacIΔ11-PAmCherry1 erm2).