pMXS-IRES-BLAST flag-CAD R1398A A1400E R1403A R1404A L1405A
(Plasmid
#188138)
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PurposeExpresses C-terminal flag-tagged human CAD R1398A A1400E R1403A R1404A L1405A in mammalian cells
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 188138 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepMXS-IRES-BLAST
- Backbone size w/o insert (bp) 5623
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Vector typeMammalian Expression, Retroviral
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Selectable markersBlasticidin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameCAD
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Alt namecarbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase
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SpeciesH. sapiens (human)
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Insert Size (bp)6702
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MutationTCC -> AGT silent mutations at nt439-441 eliminating PAM site for sgRNA target sequence 5'-ATTGGGGTCCAAGAATGGCA-3' and CG/CT/CG/CG/CT -> GC/AA/GC/GC/GC mutations at nt6114-6115/6121-6122/6129-6130/6132-6133/6135-6136 for Arg1389/Ala1400/Arg1403/Arg1404/Leu1405 -> Ala or Glu mutations of CAD
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Entrez GeneCAD (a.k.a. CDG1Z, DEE50, EIEE50, GATD4)
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Tag
/ Fusion Protein
- Flag tag (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer AGTAGACGGCATCGCAGCTTGGATA
- 3′ sequencing primer CGGAATTTACGTAGCGGCCGC (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pMXS-IRES-BLAST flag-CAD R1398A A1400E R1403A R1404A L1405A was a gift from Richard Possemato (Addgene plasmid # 188138 ; http://n2t.net/addgene:188138 ; RRID:Addgene_188138) -
For your References section:
Allosteric regulation of CAD modulates de novo pyrimidine synthesis during the cell cycle. Shin J, Mir H, Khurram MA, Fujihara KM, Dynlacht BD, Cardozo TJ, Possemato R. Nat Metab. 2023 Feb 6. doi: 10.1038/s42255-023-00735-9. 10.1038/s42255-023-00735-9 PubMed 36747088