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pLM494
(Plasmid #100539)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 100539 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pLM304
  • Backbone size w/o insert (bp) 4846
  • Total vector size (bp) 16125
  • Modifications to backbone
    Insertion of kanamycin expression cassette.
  • Vector type
    Yeast Expression, Cre/Lox, Synthetic Biology
  • Selectable markers
    URA3

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    GGPP synthase
  • Alt name
    crtE
  • Species
    X. dendrorhous
  • Insert Size (bp)
    1131
  • GenBank ID
    DQ016502.1
  • Promoter TIP1

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsaI (destroyed during cloning)
  • 3′ cloning site BsaI (destroyed during cloning)
  • 5′ sequencing primer GCTTGCCTAACTTTGTTC
  • 3′ sequencing primer GAATAGAGCAAAAGTTGGG
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    phytoene desaturase
  • Alt name
    crtI
  • Species
    X. dendrorhous
  • Insert Size (bp)
    1749
  • GenBank ID
    AY177424.1
  • Promoter PGK1

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsaI (destroyed during cloning)
  • 3′ cloning site BsaI (destroyed during cloning)
  • 5′ sequencing primer AATTGTCCGAATCGTGTG
  • 3′ sequencing primer GCGGGAAATAAGCTACAA
  • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    phytoene-beta carotene synthase
  • Alt name
    crtYB
  • Species
    X. dendrorhous
  • Insert Size (bp)
    2022
  • GenBank ID
    AY177204.1
  • Promoter TDH3

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsaI (destroyed during cloning)
  • 3′ cloning site BsaI (destroyed during cloning)
  • 5′ sequencing primer GACGGTAGGTATTGATTG
  • 3′ sequencing primer GGGACTCAAAGCGTAAAA
  • (Common Sequencing Primers)

Gene/Insert 4

  • Gene/Insert name
    HMG CoA reductase
  • Alt name
    tHMG1
  • Species
    S. cerevisiae (budding yeast)
  • Insert Size (bp)
    1575
  • GenBank ID
    M22002.1
  • Promoter ZEO1

Cloning Information for Gene/Insert 4

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsaI (destroyed during cloning)
  • 3′ cloning site BsaI (destroyed during cloning)
  • 5′ sequencing primer TCAGTTTGCTTGATAGCC
  • 3′ sequencing primer GAATAGAGCAAAAGTTGGG
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    Received from Leslie Mitchell (Boeke lab).
  • Articles Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Addgene NGS sequencing found the following mutations compared to the provided reference sequence: crtE with M176I, K346R, I347M, E353D mutations, crtI with L107F, H427Q mutations, and crtYB with A190T, E307K mutations. tHMG1 contains only residues 531-1054 compared to the NCBI reference seq [M22002.1]. These mutations/truncation do not affect plasmid function.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pLM494 was a gift from Bernd Müller-Röber (Addgene plasmid # 100539 ; http://n2t.net/addgene:100539 ; RRID:Addgene_100539)
  • For your References section:

    L-SCRaMbLE as a tool for light-controlled Cre-mediated recombination in yeast. Hochrein L, Mitchell LA, Schulz K, Messerschmidt K, Mueller-Roeber B. Nat Commun. 2018 May 22;9(1):1931. doi: 10.1038/s41467-017-02208-6. 10.1038/s41467-017-02208-6 PubMed 29789561