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

<h2>Cytosolic vectors for Chlamydomonas reinhardtii</h2> <h3>pAH04 mCherry (5536 bp)&nbsp;<a href="https://www.zotero.org/google-docs/?nUyV9D">(Molino et al., 2018)</a></h3> <ul> <li> <p>AmpR (Ampicillin Resistance):</p> </li> </ul> <p>Positioned at approximately 4000 bp, this gene confers resistance to ampicillin, enabling bacterial selection.</p> <ul> <li> <p>Amp Promoter:</p> </li> </ul> <p>Regulates the expression of the AmpR gene, facilitating its transcription and subsequent resistance to ampicillin.</p> <ul> <li> <p>f1 ori:</p> </li> </ul> <p>Positioned near 5000 bp, this origin of replication (ori) facilitates plasmid&nbsp; replication in bacterial cells, ensuring propagation of the plasmid in E. coli.</p> <ul> <li> <p>PAR1 - HSP70 Promoter + RBCS2 Promoter:</p> </li> </ul> <p>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.</p> <ul> <li> <p>mCherry:</p> </li> </ul> <p>Encodes the mCherry fluorescent protein, a red fluorescent marker commonly used for visualizing gene expression in vivo.</p> <ul> <li> <p>Ble (Bleomycin Resistance):</p> </li> </ul> <p>Provides resistance to bleomycin</p> <ul> <li> <p>Intron and UTRs (rbcS2):</p> </li> </ul> <p>Includes untranslated regions (UTRs) and introns from the rbcS2 gene, enhancing gene expression stability and processing in Chlamydomonas reinhardtii.</p> <ul> <li> <p>XbaI (93 bp) and KpnI (2750 bp):</p> </li> </ul> <p>These are key restriction enzyme sites used for cloning and linearization of the plasmid.</p> <p>&nbsp;</p> <h2>Secretion vectors for <em>Chlamydomonas pacifica</em></h2> <h3>pJPSHx1_PHL7 (5879 bp) <a href="https://www.zotero.org/google-docs/?mCtacP">(Molino, Oliver, et al., 2024)</a></h3> <ul> <li> <p>AmpR (Ampicillin Resistance):</p> </li> </ul> <p>Enables bacterial selection by conferring resistance to ampicillin.</p> <ul> <li> <p>Amp Promoter:</p> </li> </ul> <p>Drives the transcription of the AmpR gene, allowing for the maintenance of plasmids in ampicillin-containing media.</p> <ul> <li> <p>P_TUBA2 Promoter:</p> </li> </ul> <p>Promoter from the tubulin alpha-2 gene from C. pacifica that controls expression of downstream genes, including PHL7, in eukaryotic cells.</p> <ul> <li> <p>T_TUBA2</p> </li> </ul> <p>Terminator region of tubulin alpha-2 gene from C. pacifica</p> <ul> <li> <p>PHL7:</p> </li> </ul> <p>Encodes the PHL7 enzyme, a plastic-degrading enzyme, important for applications in biodegradation and recycling.</p> <ul> <li> <p>F2A Peptide:</p> </li> </ul> <p>A viral peptide sequence that allows for the co-expression and separation of proteins from a single mRNA transcript.</p> <ul> <li> <p>SP7 (Signal Peptide 7):</p> </li> </ul> <p>Directs the PHL7 protein to the secretory pathway for export from the cell.</p> <ul> <li> <p>Step Hygro aph7'' (Hygromycin Resistance):</p> </li> </ul> <p>Provides resistance to hygromycin, facilitating selection of transformed eukaryotic cells.</p> <ul> <li> <p>XbaI (56 bp) and KpnI (3056 bp):</p> </li> </ul> <p>These are key restriction enzyme sites used for cloning and linearization of the plasmid.</p> <p>&nbsp;</p> <p>&nbsp;</p> <p>&nbsp;</p>