Plasmid_Backbone
His3::EXSP
Part:BBa_K3271002
Designed by: Setti Belhouari Group: iGEM19_uOttawa (2019-10-12)
His3::EXSP pSB1K3
The prefix and suffix (EXSP) of the pSB1K3 backbone are flanked by the upstream and downstream homology sequences of the His3 locus from S. cerevisiae, respectively. To target a DNA part of interest to the His3 locus in S. cerevisiae, insert the DNA part within the prefix and the suffix using any method of your choice. Then, digest the plasmid using PmeI. This creates a DNA fragment comprised of the DNA part of interest flanked by the His3 locus upstream homology and the downstream homology at the 5' and 3' ends of the part, respectively. Transform the digest into S. cerevisiae using a lithium acetate protocol of choice.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found at 2291
Illegal suffix found at 2313 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2291
Illegal SpeI site found at 2314
Illegal PstI site found at 2328
Illegal NotI site found at 2297
Illegal NotI site found at 2321 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 2291
Illegal XhoI site found at 1026
Illegal XhoI site found at 2052 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 2291
Illegal suffix found at 2314 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 2291
Plasmid lacks a suffix.
Illegal XbaI site found at 2306
Illegal SpeI site found at 2314
Illegal PstI site found at 2328 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
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Categories
Parameters
//plasmidbackbone/assembly
biology | Saccharomyces cerevisiae |
chassis | Saccharomyces cerevisiae and Escherichia coli |
device_type | Plasmid backbone for homologous recombination in Saccharomyces cerevisiae |