pRL_CT_GI_del1
(Plasmid #98744)

• Purpose: For red/far-red light-regulated gene expression in Saccharomyces cerevisiae. Integrates into the yeast ura3-52 locus. Encodes constitutively expressed PhyBNT-synTALE-DBD and PIF3-NLS-VP64AD fusions.
• 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: pL1A-lc
• Backbone size w/o insert (bp): 4300
• Total vector size (bp): 13000
• Modifications to backbone: Additional Leu marker, additional integration cassette for the ura3-52 locus
• Vector type: Yeast Expression, Synthetic Biology
• Selectable markers: LEU2

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: phytochrome interacting factor 3
• Alt name: PIF3
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 1572
• GenBank ID: NM_100824
• Entrez Gene: PIF3 (a.k.a. AT1G09530, F14J9.19, F14J9_19, PAP3, PHOTOCURRENT 1, PHYTOCHROME INTERACTING FACTOR 3, PHYTOCHROME-ASSOCIATED PROTEIN 3, POC1, phytochrome interacting factor 3, purple acid phosphatase 3)
• Promoter: FBA1
• Tags / Fusion Proteins:
  • SV40-NLS (C terminal on insert)
  • VP64-AD (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TTCCTTCTTCTTCGCCCA
• 3′ sequencing primer: AAGAAAAGAGCCGACCAA

Gene/Insert 2

• Gene/Insert name: phytochrome B
• Alt name: PhyB
• Species: A. thaliana (mustard weed)
• Insert Size (bp): 1863
• Mutation: mutated nucleotide 576 C to A, deleted amino acids 622-1172
• GenBank ID: NM_001335612
• Entrez Gene: PHYB (a.k.a. AT2G18790, HY3, MSF3.17, MSF3_17, OOP1, OUT OF PHASE 1, PHYTOCHROME B, phytochrome B)
• Promoter: TDH3
• Tag / Fusion Protein:
  • synTALE-DBD (C terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GCAATAGCGCATCAAGAAAA
• 3′ sequencing primer: CCCTGAAATTATTCCCCTAC

Terms and Licenses

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