Ordering

This material is available to academics and nonprofits only.

Backbone

• Vector backbone
  pcDNA4/TO
  (Search Vector Database)
• Backbone manufacturer
  Invitrogen
• Backbone sizew/o insert(bp)5078
• Total vector size (bp)7446
• Modifications to backbone
  CMV-2xtetO promoter replaced with pCMV-D2i promoter. Rabbit β-globin intron and WPRE inserted before and after insert, respectively.
• Vector type
  Mammalian Expression, Synthetic Biology ; Expression regulator/reporter
• Selectable markers
  Zeocin

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
  EGFP
• Alt name
  Enhanced green fluorescent protein
• Species
  H. sapiens (human), Synthetic
• Insert Size (bp)
  1815
• 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-D2irG4kwh.PDW
• A portion of this plasmid was derived from a plasmid made by
  Mammalian codon optimized variant of eGFP gene from the pIRES2-EGFP plasmid (Clontech).Rabbit β-globin intron II-containing fragment from plasmid pcDNA6/TR (Invitrogen).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
  • Ancillary Agreement for Plasmids Containing FP Materials
• Industry Terms
  • Not Available to Industry