Expression vectors from "Description of a novel extremophile green algae, Chlamydomonas pacifica, and its potential as a biotechnology host."

Cytosolic vectors for Chlamydomonas reinhardtii

pAH04 mCherry (5536 bp) (Molino et al., 2018)

• AmpR (Ampicillin Resistance):

Positioned at approximately 4000 bp, this gene confers resistance to ampicillin, enabling bacterial selection.

• Amp Promoter:

Regulates the expression of the AmpR gene, facilitating its transcription and subsequent resistance to ampicillin.

• f1 ori:

Positioned near 5000 bp, this origin of replication (ori) facilitates plasmid  replication in bacterial cells, ensuring propagation of the plasmid in E. coli.

• PAR1 - HSP70 Promoter + RBCS2 Promoter:

A dual-promoter system that combines the heat-shock protein 70 (HSP70) and ribulose bisphosphate carboxylase small subunit 2 (RBCS2) promoters, providing strong and constitutive expression of downstream genes in Chlamydomonas.

• mCherry:

Encodes the mCherry fluorescent protein, a red fluorescent marker commonly used for visualizing gene expression in vivo.

• Ble (Bleomycin Resistance):

Provides resistance to bleomycin

• Intron and UTRs (rbcS2):

Includes untranslated regions (UTRs) and introns from the rbcS2 gene, enhancing gene expression stability and processing in Chlamydomonas reinhardtii.

• XbaI (93 bp) and KpnI (2750 bp):

These are key restriction enzyme sites used for cloning and linearization of the plasmid.

Secretion vectors for Chlamydomonas pacifica

pJPSHx1_PHL7 (5879 bp) (Molino, Oliver, et al., 2024)

• AmpR (Ampicillin Resistance):

Enables bacterial selection by conferring resistance to ampicillin.

• Amp Promoter:

Drives the transcription of the AmpR gene, allowing for the maintenance of plasmids in ampicillin-containing media.

• P_TUBA2 Promoter:

Promoter from the tubulin alpha-2 gene from C. pacifica that controls expression of downstream genes, including PHL7, in eukaryotic cells.

• T_TUBA2

Terminator region of tubulin alpha-2 gene from C. pacifica

• PHL7:

Encodes the PHL7 enzyme, a plastic-degrading enzyme, important for applications in biodegradation and recycling.

• F2A Peptide:

A viral peptide sequence that allows for the co-expression and separation of proteins from a single mRNA transcript.

• SP7 (Signal Peptide 7):

Directs the PHL7 protein to the secretory pathway for export from the cell.

• Step Hygro aph7'' (Hygromycin Resistance):

Provides resistance to hygromycin, facilitating selection of transformed eukaryotic cells.

• XbaI (56 bp) and KpnI (3056 bp):

These are key restriction enzyme sites used for cloning and linearization of the plasmid.