Description:

pYY11 is an intermediate plasmid that served as a crucial precursor in the development of the pQQC7 plastid transformation vector. This plasmid was designed and constructed to incorporate essential components and regulatory elements that were later refined and integrated into the final pQQC7 construct.

Plasmid Map of pYY11

Key Features and Role:

Foundation for pQQC7: pYY11 provided the initial backbone and key elements that were further optimized and modified to create pQQC7. It served as a testing ground for various components, ensuring their functionality and compatibility before final integration.
Cloning Flexibility: pYY11 includes multiple restriction enzyme sites, allowing for easy insertion and manipulation of genes and regulatory elements. This flexibility was crucial for the iterative design and testing phases of pQQC7.
Regulatory Elements: The plasmid contains preliminary versions of the promoters, 5' UTRs, and other regulatory sequences that were later fine-tuned in pQQC7. These elements were tested for their expression levels and stability in the chloroplast environment.
Selectable Marker and Reporter Gene: pYY11 includes the aadA selectable marker gene and the GFP reporter gene, which were used to test and validate the selection and visualization processes. These genes were driven by early versions of the psbA and Prrn promoters, respectively.
Homologous Recombination Sequences: Initial homologous recombination regions (LHRR and RHRR) were included in pYY11 to ensure proper integration into the kiwifruit chloroplast genome. These sequences were later refined and optimized in pQQC7.
Stability and Compatibility: pYY11 was designed to be stable and compatible with standard molecular biology techniques, ensuring that it could be efficiently propagated and maintained in bacterial hosts.

Summary:

pYY11 is an intermediate plasmid that played a vital role in the development of the pQQC7 plastid transformation vector. It provided the foundational structure and key components that were further refined and optimized to create the final, highly functional pQQC7 plasmid. This intermediate step was essential for ensuring the robustness and reliability of the final construct, making pYY11 a critical part of our project's success.

Usage:

Development and Testing: pYY11 can be used as a reference or starting point for researchers who are interested in the development and optimization of plastid transformation vectors.
Educational Tool: It serves as an educational tool to demonstrate the iterative process of plasmid design and the importance of intermediate steps in the development of advanced genetic constructs.

Sequence and Features

BBa_K5044311 SequenceAndFeatures