pCDNA3-2xMARS-PLD
(Plasmid #205237)

• Purpose: Expresses two repeats of PLEKAH5 aa 143-271 (K163A and R164A) fused to phospholipase D from Streptomyces sp. PMF
• Depositing Lab: Jeremy Baskin
• Publication: A phosphorylation-controlled switch confers cell cycle-dependent protein relocalization (unpublished)

Backbone

• Vector backbone: pCDNA3.1
• Backbone size w/o insert (bp): 5399
• Total vector size (bp): 8487
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: 2xPLEKHA5(143-271)-K163A/R164A-HA-nGFP
• Alt name: PEPP2
• Species: H. sapiens (human)
• Insert Size (bp): 834
• Mutation: amino acids 143-271 of PLEKHA5 (GenBank reference sequence NM_001256470.2) with K163A and R164A mutations
• GenBank ID: NM_001256470.2
• Entrez Gene: PLEKHA5 (a.k.a. PEPP-2, PEPP2)
• Promoter: CMV
• Tags / Fusion Proteins:
  • Nuclear Export Sequence (C terminal on insert)
  • mCherry (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: mCherry reverse

Gene/Insert 2

• Gene/Insert name: phospholipase D (PLD)
• Species: Streptomyces sp. PMF
• GenBank ID: UniProtKB P84147
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: mCherry forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pCDNA3-2xMARS-PLDwt.gbk

Terms and Licenses

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