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A memory switch for plant synthetic biology based on the phage varphiC31 integration system.

Bernabe-Orts JM, Quijano-Rubio A, Vazquez-Vilar M, Mancheno-Bonillo J, Moles-Casas V, Selma S, Gianoglio S, Granell A, Orzaez D
Nucleic Acids Res. 2020 Apr 6;48(6):3379-3394. doi: 10.1093/nar/gkaa104. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
160557pUPD 5'UTR:YFP:Tnos (GB1483)Part 3/3 for assembly of site-specific recombination based Registers. Composed of the 5'UTR sequence of 35S promoter, coding sequence of YFP and NOS terminator (5'UTR:YFP:TNos)
160577pUPD2 PhiC31 attP:TMtb:P35S:attB (GB1494)35S promoter sequence and the Mtb terminator flanked by two opposing PhiC31 site-specific recombination sites (attP and attB)
160578pUPD2 PhiC31 integrase (GB1496)Streptomyces phage PhiC31 integrase, complete CDS
160579pUPD2 RDF ( GB1498)Streptomyces phage PhiC31-encoded recombination directionality factor (RDF) (PhiC31 gp3)
160580pUPD2 TNos:Luc:5'UTR (GB1499)Reversed sequences of the NOS terminator, Firefly Luciferase CDS and 5' UTR of 35S promoter
160581pUPD2 Assembly Register 3/3 Part - 5'UTR:Luc:Tnos (GB1500)Part 3/3 for assembly of site-specific recombination based Registers. Composed of the 5'UTR sequence of 35S promoter, coding sequence of Luc and NOS terminator
160582pUPD2 PhiC31 RL (attR:P35S:TMtb:attL) (GB1506)35S promoter inverted sequence and the Mtb terminator flanked by two opposing PhiC31 site-specific recombination sites (attR and attL)
160583pUPD2 Register 1/3 Tnos:YFP:5'UTR (GB1507)Part 1/3 for assembly of site-specific recombination based Registers. Composed of the reversed sequences of the TNos terminator,YFP CDS and 5' UTR of 35S promoter (TNos:YFP:5'UTR)
160586pUPD2 PhiC31 integrase (GB1561)Streptomyces phage PhiC31 integrase, adapted for C-terminal fusions
160604pUPD2 linker-RDF (GB2892)RDF sequence for C-terminal fusion with PhiC31 recombinase, together with a 18-nt linker sequence
160613pEGB3 alpha2 P35S:RDF:T35S (GB1508)TU for the constitutive expression of Streptomyces phage PhiC31-encoded recombination directionality factor (RDF) (PhiC31 gp3)
160614pEGB3 alpha2 OplexA:mini35S:PhiC31int:Tnos (GB1529)TU for the regulated expression of the PhiC31 phage integrase gene driven by a synthetic promoter consisting of the LexA operator fused to the minimal 35S promoter.
160615pEGB_3alpha1 OplacI:mini35S:phiC31:Tnos (GB1530)TU for the regulated expression of the PhiC31 phage integrase gene driven by a synthetic promoter consisting of the LacI operator fused to the minimal 35S promoter.
160616pEGB3 alpha1 Pnos:PhiC31int:Tnos (GB1531)TU for the constitutive expression of Streptomyces phage PhiC31 integrase.
160617pEGB_3alpha2 OplacI:mini35S:RDF:Tnos (GB1534)TU for the regulated expression of the PhiC31 phage recombination directionality factor (RDF) gene driven by a synthetic promoter consisting of the LacI operator fused to the minimal 35S promoter.
160619pDGB3_alpha2: Pnos:ER:lexABD:GAL4AD:T35s-OplexA:mini35S:phi31:Tnos-Pnos:GR:LacIBD:Gal4AD:Tnos-OplacI:mini35S:RDF:Tnos (GB1679)Module for estradiol-inducible expression of the PhiC31 integrase gene and dexamethasone -inducible expression of PhiC31 phage recombination directionality factor (RDF) gene.
160633pEGB3 alpha1 P35S:PhiC31 integrase-RDF:T35S (GB2893)TU for the constitutive expression of the PhiC31 integrase, C-terminally fused with its reversion factor (RDF)
160641pDGB3_Omega1_Pnos:ER:lexABD:GAL4AD:T35s-OplexA:mini35S:phi31:Tnos (GB1677)Module for estradiol-inducible exp of the PhiC31 integrase gene. Includes TU for constitutive exp of an estradiol-inducible transcription activator and TU for estradiol-inducible exp of PhiC31.
160642pDGB3_Omega2_Pnos:GR:LacIBD:Gal4AD:Tnos-OplacI:mini35S:RDF:Tnos (GB1678)Module for the dexamethasone -inducible expression of PhiC31 phage recombination directionality factor (RDF) gene.

Antibodies from Article