pMDS1
(Plasmid #239460)

• Purpose: (Empty Backbone) For simultaneous monitoring of both transcription and translation of genes in plant tissues: C-terminus fusion
• Depositing Lab: Miguel de Lucas
• Publication: Banda et al Sci. Adv., 11, eadw9153 (2025)

Backbone

• Vector backbone: pMCY2
• Modifications to backbone: We have incorporated a module that allows the visualisation of transcription and translation in one go. pMDS1 provides the following C-terminal fusion VENUS:3xHA:2A:mTurqN7
• Vector type: Bacterial Expression, Plant Expression
• Selectable markers: Plasma membrane mCherry marker for seed selection
• Tag / Fusion Protein:
  • VENUS:3xHA:2A:mTurqN7 (C terminal on insert)

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin and Streptomycin, 50 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Successful cloning can be tested via Blue-White selection
• Copy number: High Copy

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• Supplemental Documents:
  • pMDS1_Addgene.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

For cloning a promoter + gene fragment (without stop codon) via Gibson Assembly, we recommend the following approach:

Primer design:
Forward primer: 5’GATCGGGGCGCGCCCTCGAG+PROMOTER SPECIFIC PRIMER 3’
Reverse primer:  5’ CAGAACCACCACCTCCGGATCC+GENE SPECIFIC PRIMER W/O STOP CODON 3'

Digest pMDS1 plasmid with SAP I enzyme
Clean up

Perform Gibson assembly reaction
Transform E. coli (DH5alpha)
Select on LB + Sp + Sm