Ordering

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Backbone

• Vector backbone
  pcDNA6/TR
  (Search Vector Database)
• Backbone manufacturer
  Invitrogen
• Backbone sizew/o insert(bp)6662
• Total vector size (bp)6823
• Modifications to backbone
  CMV promoter replaced with pCMV-D2i promoter. Rabbit β-globin intron and TetR replaced with new Rabbit β-globin intron-mCherry-WPRE fragment.
• Vector type
  Mammalian Expression, Synthetic Biology ; Expression regulator/reporter
• Selectable markers
  Blasticidin

Growth in Bacteria

• Bacterial Resistance(s)
  Ampicillin
• Growth Temperature
  37°C
• Growth Strain(s)
  XL10 Gold
• Copy number
  Unknown

Gene/Insert1

• Gene/Insert name
  pCMV-D2i promoter
• Alt name
  Modified CMV-based synthetic promoter containing two tetO2 sites
• Species
  Synthetic
• Insert Size (bp)
  672
• Mutation
  Initiator motif (Inr) displaced relative to pCMV-2xtetO (returned to its natural position).Two tetO2 sites flanking Inr motif.
• PromoterpCMV-D2i

Cloning Informationfor Gene/Insert 1

• Cloning methodRestriction Enzyme
• 5′ cloning siteSpeI(not destroyed)
• 3′ cloning siteAflII(not destroyed)
• 5′ sequencing primerAmp-R
• 3′ sequencing primerBglob-intron-R (TTTGCCCCCTCCATATAACA)
(Common Sequencing Primers)

Gene/Insert2

• Gene/Insert name
  mCherry
• Alt name
  monomeric dsRed derivative
• Species
  H. sapiens (human), Synthetic
• Insert Size (bp)
  1806
• Mutation
  Region around the ATG translation start codon converted to the consensus Kozak sequence; sequence optimized for mammalian codon bias
• Tags/ Fusion Proteins
  • Rabbit β-globin intron II (N terminal on insert)
  • WPRE (C terminal on insert)

Cloning Informationfor Gene/Insert 2

• Cloning methodRestriction Enzyme
• 5′ cloning siteBamHI(not destroyed)
• 3′ cloning siteXhoI(not destroyed)
• 5′ sequencing primerBglob-intron-F (ctggtcatcatcctgccttt); CMV-F
• 3′ sequencing primerWPRE-R; BGH-rev
(Common Sequencing Primers)

Resource Information

• Supplemental Documents
  • pDN-D2irC6kwh.PDW
• A portion of this plasmid was derived from a plasmid made by
  Fragment containing the mCherry gene from the pTRE-Dual2 plasmid (Clontech).Fragment containing WPRE from plasmid pLVX-DD-tdTomato (Clontech).Modified CMV-based synthetic promoter pCMV-D2i synthesized de novo (Bio Basic Inc., Markham, Ontario, Canada).
• Terms and Licenses
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms
  • Not Available to Industry