pucKan_ARS_pgkZeo_ncx1_mCherry_2TA_Puro_ARS
(Plasmid #163755)

• Purpose: piggybac transposon encoding human Ncx1 gene promoter-driven mCherry and puromycin-acetyltransferase for labelling and selection of pluripotent stem cell-derived cardiomyocytes
• Depositing Lab: Kurt Pfannkuche
• Publication: Hamad et al Methods Mol Biol. 2021 Oct 19. doi: 10.1007/7651_2021_395.

Backbone

• Vector backbone: pucKan
• Backbone manufacturer: GeneScript
• Backbone size w/o insert (bp): 2562
• Total vector size (bp): 9691
• Vector type: Mammalian Expression ; piccybac transposon
• Selectable markers: Zeocin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mCherry
• Species: Discosoma
• Promoter: human NCX1
• Tag / Fusion Protein:
  • puromycin-N-acetyltransferase (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: NN
• 3′ sequencing primer: NN

Resource Information

• Supplemental Documents:
  • pucKan_ARS_pgkZeo_ncx1_mCherry_2TA_Puro_ARS.gbk
• A portion of this plasmid was derived from a plasmid made by: This gene was generated by gene synthesis. Sequences elements were obtained from previously published material. (1) piggybac terminal repeats were taken from pXL-BacII (Xu Li and Malcolm J. Fraser). (2) mCherry-T2A-Puro_WPRE element was taken from addgene #72264 (3) the construct is flanked 5 and 3 prime with double copies of ARS barriers. The Ars insulator sequence was kindly provided by Dr. Masao Matsuoka. (4) a sequence for Zeocin resistance was obtained from a commercial vector. (5) human pgk promoter was extracted from addgene #12252

Terms and Licenses

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