pAMVe
(Plasmid #249494)

• Purpose: Expresses mVenus – Monodirectional Chloroplast Expression Vector (psbH Rescue) in Chlamydomonas Reinhardtii photosynthetically deficient strain CC4388
• Depositing Lab: Isabel Desgagné-Penix
• Publication: Engineering bidirectional chloroplast promoters for tunable co-expression of multiple genes in microalgae (unpublished)

Backbone

• Vector backbone: pASAPI
• Backbone manufacturer: Chlamydomonas Resource Center; http://www.chlamycollection.org
• Backbone size w/o insert (bp): 6295
• Total vector size (bp): 7031
• Modifications to backbone: The psbD/ 5′ UTR, a light-activated chloroplast promoter enabling expression of the endogenous psbD gene and the psbA/ 3′ UTR of the psbA gene were added
• Vector type: Bacterial Expression, Synthetic Biology ; Chlamydomonas reinhardtii chloroplast genome
• Selectable markers: To be use in Chlamydomonas reinhardtii strain CC4388, which is photosynthetically deficient

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: NEB Stable E. coli strain can also be used
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mVenus (yellow/green fluorescence)
• Alt name: mVenus
• Species: Aequorea victoria
• Insert Size (bp): 719
• Mutation: Changed of F46L, F64L, M153T, V163A, S175G
• GenBank ID: GenBank: AAZ65844 GenBank: AAZ65844
• Promoter: The psbD/ 5′ UTR, a light-activated chloroplast promoter enabling expression of the endogenous psbD gene
• Tags / Fusion Proteins:
  • No
  • No

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: complete plasmid sequencing
• 3′ sequencing primer: complete plasmid sequencing

Terms and Licenses

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

Depositor Comments

The plasmid pAMVe is designed for expression of one transgene in the chloroplast of Chlamydomonas reinhardtii using the monodirectional promoter psbD/ 5′ UTR. This vector is intended for use in the C. reinhardtii strain CC4388, which is photosynthetically deficient due to disruption of the psbH gene, encoding a small subunit of Photosystem II. Correct integration of the plasmid into the chloroplast genome restores psbH function and rescues photosynthesis. Selection is therefore performed by photoautotrophic growth, without the need for antibiotic selection.