pRL_GI_PCB28
(Plasmid #98745)

• Purpose: For phycocyanobilin production in Saccharomyces cerevisiae. Contains integrative cassette for the yeast his1-Δ200 locus. Encodes constitutively expressed HY1 and PcyA.
• Depositing Lab: Bernd Müller-Röber
• Publication: Hochrein et al Nucleic Acids Res. 2017 Sep 6;45(15):9193-9205. doi: 10.1093/nar/gkx610.

Backbone

• Vector backbone: pL0C_4-R
• Backbone size w/o insert (bp): 5864
• Total vector size (bp): 9733
• Modifications to backbone: Insertion of a TRP marker, Insertion of a genome integration cassette for the his1-Δ200 locus
• Vector type: Yeast Expression, Synthetic Biology
• Selectable markers: TRP1, URA3

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: heme oxygenase 1
• Alt name: HY1
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 849
• GenBank ID: AB021858
• Entrez Gene: TED4 (a.k.a. AT2G26670, ARABIDOPSIS THALIANA HEME OXYGENASE 1, ATHO1, F18A8.4, F18A8_4, GENOMES UNCOUPLED 2, GUN2, HEME OXYGENASE, HEME OXYGENASE 1, HEME OXYGENASE 6, HO1, HY1, HY6, PLASTID HEME OXYGENASE, REVERSAL OF THE DET PHENOTYPE 4)
• Promoter: PGK1

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CGTAAAGGATGGGGAAAGAGA
• 3′ sequencing primer: GTTTAGTAGAACCTCGTG

Gene/Insert 2

• Gene/Insert name: Phycocyanobilin:ferredoxin oxidoreductase
• Alt name: PcyA
• Species: Synechocystis sp. PCC 6803
• Insert Size (bp): 747
• Mutation: Codon optimized for Saccharomyces cerevisiae
• Promoter: ADH1m

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CCCTTTCTTCCTTGTTTC
• 3′ sequencing primer: GGTGTGGTCAATAAGAGC

Resource Information

• Supplemental Documents:
  • pRL_GI_PCB28.gbk

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry