pGreenIIM DR5v2-ntdTomato/DR5-n3GFP
(Plasmid #61628)

• Purpose: Expresses nuclear tdTomato from auxin-dependent DR5v2 (TGTCGG) promoter and nuclear 3xGFP from the DR5 (TGTCTC) promoter
• Depositing Lab: Dolf Weijers
• Publication: Liao et al Nat Methods. 2015 Mar;12(3):207-10. doi: 10.1038/nmeth.3279. Epub 2015 Feb 2.

Backbone

• Vector backbone: pGreenII
• Backbone size w/o insert (bp): 2580
• Total vector size (bp): 9692
• Vector type: Plant Expression
• Selectable markers: Methothrexate

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: ntdTomato
• Species: Synthetic
• Insert Size (bp): 1518
• GenBank ID: KJ081243.1
• Promoter: Auxin-dependent DR5v2 promoter containing 9 reverse TGTCGG sites and -46 35S promoter

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (unknown if destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: M13_pUC_fwd

Gene/Insert 2

• Gene/Insert name: n2xEGFP
• Species: Synthetic
• Insert Size (bp): 2100
• Mutation: 2 copies of EGFP with N-terminal SV40 NLS
• GenBank ID: AF302837
• Promoter: Auxin-dependent DR5 promoter containing 9 reverse TGTCTC sites and -46 35S promoter

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: dsRed1_C_primer

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Test
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please note that Addgene NGS results identified as stop codon after the second EGFP ORF. This stop codon was present in the original, published version of the plasmid, such that the insert is n2GFP rather than n3GFP. The plasmid functions as described in the associated publication. Co-expression with the helper plasmid pSOUP is suggested for expression in Agrobacterium.

The diagnostic digest indicates this plasmid may be bigger than the assembled sequence suggests.