Plasmid_Backbone

Part:BBa_K4365023

Designed by: Giorgio Gilioli   Group: iGEM22_TU_Dresden   (2022-09-30)
Revision as of 08:23, 12 October 2022 by Svetlana ia (Talk | contribs) (→‎Usage and Biology)

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pSB1KY-TRP1

The pSB1KY-TRP1 is a shuttle plasmid backbone suitable for the iGEM RFC1000 assembly in Saccharomyces cerevisiae with TRP1 auxotrophy. It was adapted from pSB1K plasmid and optimized to be expressed in Escherichia coli cells and S. cerevisiae with trp1 mutations.


Collection of shuttle vector backbones optimized for Saccharomyces cerevisiae
Backbone ID Selection marker Marker sequence source
BBa_K4365022 URA3 [http://freegenes.github.io/genes/BBF10K_000474.html Open Yeast Collection ScURA3-marker]
BBa_K4365023 TRP1 [http://freegenes.github.io/genes/BBF10K_000473.html Open Yeast Collection ScTRP1-marker]
BBa_K4365024 HIS3 [http://freegenes.github.io/genes/BBF10K_000468.html Open Yeast Collection ScHIS3-marker]


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 465
    Illegal SpeI site found at 1473
    Illegal PstI site found at 4
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal NheI site found at 2909
    Illegal SpeI site found at 1473
    Illegal PstI site found at 4
    Illegal NotI site found at 3112
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BamHI site found at 100
    Illegal XhoI site found at 4040
    Illegal XhoI site found at 5066
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 465
    Illegal SpeI site found at 1473
    Illegal PstI site found at 4
  • 25
    INCOMPATIBLE WITH RFC[25]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal XbaI site found at 465
    Illegal SpeI site found at 1473
    Illegal PstI site found at 4
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.


Usage and Biology

TRP1 codes for phosphoribosylanthranilate isomerase that catalyzes the third step in tryptophan biosynthesis [1]. It complements trp1 mutations allowing yeast to grow on media with no tryptophan supplements.

The pSB1KY-HIS3 was assembled similarly to pSB1KY-URA3.

  1. ↑ https://www.yeastgenome.org/locus/s000002414
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Categories
Parameters
None