pMXS-IRES-BLAST flag-CAD G-Loop1
(Plasmid #188128)

• Purpose: Expresses C-terminal flag-tagged human CAD G-Loop1 in mammalian cells
• Depositing Lab: Richard Possemato
• Publication: Shin et al Nat Metab. 2023 Feb 6. doi: 10.1038/s42255-023-00735-9.

Backbone

• Vector backbone: pMXS-IRES-BLAST
• Backbone size w/o insert (bp): 5623
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Blasticidin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: CAD
• Alt name: carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase
• Species: H. sapiens (human)
• Insert Size (bp): 6702
• Mutation: TCC -> AGT silent mutations at nt439-441 eliminating PAM site for sgRNA target sequence 5'-ATTGGGGTCCAAGAATGGCA-3' and TTTGAG...CCACCA -> GGGGGC...GGTGGA substitution at nt4096-4128 for serial glycine mutations of CAD residues F1366-P1376
• Entrez Gene: CAD (a.k.a. CDG1Z, DEE50, EIEE50, GATD4)
• Tag / Fusion Protein:
  • Flag tag (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AGTAGACGGCATCGCAGCTTGGATA
• 3′ sequencing primer: CGGAATTTACGTAGCGGCCGC

Terms and Licenses

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