Team:Valencia UPV/Parts
Know our parts Our team sent 14 Parts to the Registry: BBa_K1742000 - AsLOVpep AsLOVpep is a protein composed by the second LOV (light-oxigen-voltage) domain of Avena sativa phototropin and a C-terminus peptide tag for ePDZ recognition. This part was used to complete a blue light-induced switch (along with BBa_K1470005) present in the first and second modules of our biological circuit. BBa_K1742001 - Enterotoxin LTB The LTB is a heat-labile enterotoxin beta subunit from Escherichia coli. This DNA sequence underwent codon usage optimization for Nicotiana tabacum. LTB protein was used to test its expression levels in N. benthamiana leaves. BBa_K1742002 - Dronpa 145N Dronpa 145N is a green fluorescence protein that contains a K145N substitution. It is switched off upon illumination with cyan light and switched on with violet light. It is also capable to form heterodimers with Dronpa K (BBa_K1742003) upon violet light exposure. This DNA sequence underwent codon usage optimization for Arabidopsis thaliana. This part was used to test its potential as a transgene expression system. BBa_K1742003 - Dronpa 145K Dronpa 145K is the wild type protein used as a template to obtain Dronpa 145N (BBa_K1742002). This part was used to test its potential as a transgene expression system BBa_K1742004 - PhiC31 Plant codon optimized PhiC31 is a site-specific serine recombinase derived from the PhiC31 phage. The enzyme recognizes two different attachments sites called also attB and attP, and also excise a sequence flanked with attB and attP sites. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana. We used PhiC31 as excisionase by flanking a T35S terminator with the recognition sites. BBa_K1742005 - BxBI integrase Bxb1 is a serine recombinase protein from Mycobateriophage Bxb1’s gp35 gen. This integrase is able to recognize two different sites, attP and attB. Depending on the position and sense of these sites bxb1 is able of excising or inverting. We used bxb1 as excisionase by flanking a T35S terminator with the recognition sites. BBa_K1742006 - BxBI Reporter (attB:T35S:attP:OmegaUTR) The part consists of Cauliflower Mosaic Virus T35S terminator flanked by the BxbI recognition sites attB and attP. It also contains an Omega UTR region. This part was used to test BxbI activity in N. benthamiana plants. BBa_K1742007 - LTF Lactoferrin This part codifies the complete Homo sapiens lactoferrin CDS. BBa_K1742008
- PhiC31 Reporter (attP:T35S:aatB:OmegaUTR) The part consists of Cauliflower Mosaic Virus T35S terminator flanked by the BxbI recognition sites attP and attB. It also contains an Omega UTR region. This part was used to test PhiC31 activity in N. benthamiana plants. BBa_K1742009 - 35S:BxbI:T35S - 35S:ReporterBxBI::GFP:T35S This device consists of two transcriptional unit modules: the BxbI integrase (BBa_K1742005) driven by the constitutive promoter P35S and the reporter element (BBa_K1742006) that contains the recognition sites flanking a T35S terminator and a GFP protein as a gene marker. When BxbI is expressed, it recognizes the attB and attP sites, excising the terminator and provoking the transcription of GFP. BBa_K1742010 - 35S:LexABD-DronpaK:T35S - 35S:DronpaN-VP16:T35S This device for plants consists of two modules: A Dronpa K fused to the DNA binding domain LexA, and a second transcriptional unit that consists of Dronpa N fused to the transcriptional activator VP16. Upon illumination with violet light, it is produced a heterodimerizacion between Dronpa K and N, leading to the expression of a gene of interest driven by LexA operator and a minimal promoter mini35S. This part was used to test the light-induced toggle switch functionality. BBa_K1742011 - 35S:LacIBDBD-DronpaK:T35S - 35S:DronpaN-VP16:T35S This device for plants consists of two modules: A Dronpa K fused to the DNA binding domain LacI, and a second transcriptional unit that consists of Dronpa N fused to the transcriptional activator VP16. Upon illumination with violet light, it is produced a heterodimerizacion between Dronpa K and N, leading to the expression of a gene of interest driven by LacI operator and a minimal promoter mini35S. This part was used to test the light-induced toggle switch functionality. BBa_K1742012 - 35S:Gal4BD-DronpaK:T35S - 35S:DronpaN-VP16:T35S This device for plants consists of two modules: A Dronpa K fused to the DNA binding domain Gal4, and a second transcriptional unit that consists of Dronpa N fused to the transcriptional activator VP16. Upon illumination with violet light, it is produced a heterodimerizacion between Dronpa K and N, leading to the expression of a gene of interest driven by UAS operator and a minimal promoter mini35S. This part was used to test the light-induced toggle switch functionality. BBa_K1742013 - 35S:PhiC31:T35S - 35S:ReporterPhiC31::GFP:T35S This device consists of two transcriptional unit modules: the PhiC31 integrase (BBa_K1742004) driven by the constitutive promoter P35S and the reporter element (BBa_K1742008) that contains the recognition sites flanking a T35S terminator and a GFP protein as a gene marker. When PhiC31 is expressed, it recognizes the attP and attB sites, excising the terminator and provoking the transcription of GFP. Valencia UPV team did also improve parts that already existed in the Registry:
Parts
Part Number Part Name Backbone Type Length BBa_K1742000
AsLOVpep pSB1C3 Coding 456 BBa_K1742001 Enterotoxin LTB pSB1C3 Coding 414 BBa_K1742002 Dronpa 145N pSB1C3 Coding 693 BBa_K1742003 Dronpa 145K pSB1C3 Coding 741 BBa_K1742004 PhiC31 Plant codon optimized pSB1C3 Coding 1836 BBa_K1742005 BxBI integrase pSB1C3 Coding 1693 BBa_K1742006 BxBI Reporter (attB:T35S:attP:OmegaUTR) pSB1C3 Reporter 397 BBa_K1742007 LTF Lactoferrin pSB1C3 Coding 2079 BBa_K1742008
PhiC31 Reporter (attP:T35S:aatB:OmegaUTR) pSB1C3 Reporter 412 BBa_K1742009 35S:BxbI:T35S - 35S:ReporterBxBI::GFP:T35S pSB1C3 Device 2964 BBa_K1742010
35S:LexABD-DronpaK:T35S - 35S:DronpaN-VP16:T35S pSB1C3 Device 3176 BBa_K1742011 35S:LacIBDBD-DronpaK:T35S - 35S:DronpaN-VP16:T35S pSB1C3 Device 2592 BBa_K1742012 35S:Gal4BD-DronpaK:T35S - 35S:DronpaN-VP16:T35S pSB1C3 Device 3610 BBa_K1742013 35S:PhiC31:T35S - 35S:ReporterPhiC31::GFP:T35S pSB1C3 Device 5508
Improved parts