p28-2iDMD-puro
(Plasmid #88895)

• Purpose: 28°C full-length DMD luc-puro-red plasmid. Co-expresses human DMD, firefly luciferase, puroR and mCherry in mammalian cells. Plasmid carries phiC31 and Bxb1 attB sites, but *cannot* be grown at 37°C.
• Depositing Lab: Michele Calos
• Publication: Farruggio et al Biotechnol J. 2017 Jan 31. doi: 10.1002/biot.201600477.

Backbone

• Vector backbone: p28
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 25 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH10B
• Growth instructions: We recommend growth at 28°C. Depositor recommends DH10B, XL1 blue or NEB's DH10-beta for growth. DH5alpha should not be used as it does not yield good transformation results
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: Dystrophin
• Alt name: DMD
• Species: H. sapiens (human)
• Insert Size (bp): 11262
• Entrez Gene: DMD (a.k.a. BMD, CMD3B, DXS142, DXS164, DXS206, DXS230, DXS239, DXS268, DXS269, DXS270, DXS272, MRX85)

Gene/Insert 2

• Gene/Insert name: luciferase
• Alt name: luc
• Species: Firefly
• Insert Size (bp): 1647

Gene/Insert 3

• Gene/Insert name: puromycin resistance enzyme
• Alt name: puroR
• Insert Size (bp): 594
• Mutation: codon optimized

Gene/Insert 4

• Gene/Insert name: mCherry
• Alt name: red
• Insert Size (bp): 705

Resource Information

• Supplemental Documents:
  • p28-2iDMD-puro.gbk
• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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

Depositor Comments

Plasmid *cannot* be grown at 37°C