pSB-CMV-MCS-Puro cPLA2-mK2-P2A-EGFP-LaminB1
(Plasmid #162566)

• Purpose: Used for the dual color labeling of cPLA2 and LaminB1 expression.
• Depositing Lab: Philipp Niethammer
• Publication: Enyedi et al Cell. 2016 May 19;165(5):1160-1170. doi: 10.1016/j.cell.2016.04.016.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5428
• Total vector size (bp): 11236
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: cytosolic phospholipase a2
• Alt name: cPLA2
• Alt name: pla2g4aa
• Species: D. rerio (zebrafish)
• Insert Size (bp): 2962
• GenBank ID: 30554
• Entrez Gene: pla2g4aa (a.k.a. PLA2G4A, cpla2, pla2g4)
• Promoter: CMV
• Tags / Fusion Proteins:
  • mKate2 (C terminal on insert)
  • P2A (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV
• 3′ sequencing primer: BGHR

Gene/Insert 2

• Gene/Insert name: Lamin B1
• Alt name: LMNB1
• Species: D. rerio (zebrafish)
• Insert Size (bp): 1767
• GenBank ID: 195816
• Entrez Gene: lmnb1 (a.k.a. fc06g01, wu:fc06g01)
• Promoter: SV40
• Tag / Fusion Protein:
  • EGFP (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV
• 3′ sequencing primer: BGHR

Resource Information

• Supplemental Documents:
  • pSB-CMV-MCS-Puro cPLA2-mKate2-P2A-EGFP-LMNB1.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry