pDB2-hPER1-mScarlet-I-CD4-bla
(Plasmid #189980)

• Purpose: Donor Vector to integrate mScarlet-I C-terminal into the human PER1 Locus to express it as a fusion protein
• Depositing Lab: Achim Kramer
• Publication: Gabriel et al Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2404738121. doi: 10.1073/pnas.2404738121. Epub 2024 Aug 14.

Backbone

• Vector backbone: pDB2, derived from pUC57
• Backbone size w/o insert (bp): 8315
• Total vector size (bp): 9008
• Modifications to backbone: regions homologous to the hPER1 genomic region (surrounding STOP codon) flank insert + floxed positive selection cassette (mCerulean-P2A-blaR). Silent mutations to the hPER1 genomic sequence were inserted to avoid donor vector targeting by Cas9/sgRNA. Negative Selection cassette expresses hCD4. Should be used together with Cas9/sgRNA plasmid #189987 or #189988
• Vector type: CRISPR
• Selectable markers: Blasticidin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: mScarlet-I
• Species: Synthetic
• Insert Size (bp): 693
• Tag / Fusion Protein:
  • 3xFlag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: MluI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: --
• 3′ sequencing primer: --

Resource Information

• Supplemental Documents:
  • pDB2.0 Per1-5'+3' Scarlet.gb
• A portion of this plasmid was derived from a plasmid made by: hCD4 was cloned from Addgene #35712, mScarlet was cloned from #98839

Terms and Licenses

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

Depositor Comments

Please visit https://www.biorxiv.org/content/10.1101/2024.02.06.579141v1 for bioRxiv preprint.