Part:BBa_K5371003:Design
pRS42H
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 836
Illegal XbaI site found at 7581
Illegal PstI site found at 926 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 836
Illegal PstI site found at 926 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 836
Illegal BamHI site found at 3161
Illegal XhoI site found at 3887 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 836
Illegal XbaI site found at 7581
Illegal PstI site found at 926 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 836
Illegal XbaI site found at 7581
Illegal PstI site found at 926
Illegal NgoMIV site found at 2415 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 2915
Illegal BsaI.rc site found at 5751
Illegal SapI site found at 4668
Design Notes
When designing the pRS42H plasmid backbone, several factors were taken into account:
Selection Markers: The inclusion of both hygromycin and ampicillin resistance markers allows for easy selection in both yeast and bacterial hosts. High-Copy Maintenance: The 2-micron origin of replication ensures high-copy maintenance of the plasmid in yeast cells, which is crucial for achieving high levels of gene expression. Cloning Sites: Multiple cloning sites were incorporated to facilitate the insertion of various genes and regulatory elements. Compatibility: The plasmid was designed to be compatible with standard yeast and bacterial cloning techniques, making it versatile and easy to use in different experimental setups.
Source
The pRS42H plasmid backbone is derived from the yeast Saccharomyces cerevisiae. It was constructed by integrating various functional elements, including the 2-micron origin of replication, hygromycin resistance gene, and ampicillin resistance gene, into a single vector. We would like to express our sincere gratitude to Professor Lian Jiazhang from Zhejiang University for providing the pRS42H plasmid backbone. His generous support and valuable resources have been instrumental in the success of our project.