pDB-hPER2-mClover3-CD4-bla
(Plasmid #179448)

• Purpose: Donor vector to knock in mClover3 N-terminal to human PER2 gene
• Depositing Lab: Achim Kramer
• Publication: Gabriel et al Nat Commun. 2021 Jun 18;12(1):3796. doi: 10.1038/s41467-021-24086-9.

Backbone

• Vector backbone: pDB, derived from pUC57
• Backbone manufacturer: none
• Backbone size w/o insert (bp): 8200
• Total vector size (bp): 9013
• Modifications to backbone: regions homologous to the hPER2 genomic region (surrounding STOP codon) flank insert+positive selection cassette (mCerulean-P2A-blaR). Silent mutations to the hPER2 genomic sequence were inserted to avoid donor vector targeting by Cas9/sgRNA. Negative Selection cassette expresses hCD4. Should be used together with Cas9/sgRNA plasmids #179454, 179455, 179456
• 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: mClover3
• Species: Aequorea victoria
• Insert Size (bp): 714
• Promoter: None
• Tag / Fusion Protein:
  • His/Flag (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pDB-hPER2-mClover3-CD4-bla.gb
• A portion of this plasmid was derived from a plasmid made by: hCD4 was cloned from Addgene #35712, mClover3 was cloned from #74252

Terms and Licenses

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