CRY2 and CIB1 are extracted from Arabidopsis thaliana. tCas9 is from GE Share company. VP64 is synthetic construct.
ihpRNA construct against Arabidopsis Bet v 1 allergen
Intron containing hairpin RNA construct for targeting knockdown of the Bet v 1 allergen homolog in Arabidopsis. Assembled with PDK intron from pHANNIBAL.
Intermediate parts PCRed from Arabidopsis genomic DNA and pHANNIBAL vector. Assembled using Silver Standard 21
A constitutive plant actin promoter followed by two copies of the Lac operator. This allows control of promoter activity by LacI.
pORE Gus Reporter Series Vector with BiOBrick MCS (Kan resistance)
Vector for transformation of plants. Cloning in E. coli, transfer to Agrobacterium, integrates into plant genome. Kan resistance marker in bacteria, confers Basta resistance in plants. Has a constitutive plant promoter and in-frame Gus reporter. Conforms to Silver BioBrick Assembly Standard 21.
Modified vector pORE_R from the Arabidopsis Information Resource
amiRNA Construct for lycopene beta cyclase (LYC)
Plant artificial microRNA construct for targeted knockdown of LYC.
MicroRNA targeting sequences designed with the Web MicroRNA Designer: http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Designer;project=stdwmd
Modified RS300 plasmid: http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Home;project=stdwmd
Expa4 plant secretion signal, localizes to extracellular space
Coding region for first 20 aminoacids of plant protein Exansin4. Secretion signal for plants (derived from A. thaliana). When fused to a protein, directs the protein to extracellular space.
Sequence was optimized for Escherichia coli and Arabidopsis thaliana codon usage.
Derived from Expansin4 from Arabidopsis thaliana(NP_181500).
Brazzein is a sweet-tasting protein that is found in the Western African fruit, the Oubli (Pentadiplandra brazzeana). The protein consists of a 54 amino acid sequence. It is sweeter than sugar and is used as an alternative low calorie sweetener.
Synthesis was performed by Mr. Gene, optimized for Arabidopsis expression. Based on Genbank sequence ACK76425.1.
Arabidopsis optimized Brazzein with StrepII N-terminus
Arabidopsis optimized Brazzein (BBa_K382020) with a StrepII tag for western blot analysis.
BBa_K382020 with biobricked strepII tag obtained from the Silver Laboratory.
Prokaryotic Cryptochrome 2 (CRY2) ( a blue light stimulated photoreceptor)
Cryptochrome 2 (CRY2) is a blue light stimulated photoreceptor, when exposed to blue light, it would interact withthe N-terminal fragment of CIB1 (CIBN) . This part is full-length CRY2 CDS. Optogenetic systems enable precise spatial and temporal control of cell behavior. A light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light.This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive tCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes.
mutant EcoR 1 : TGTTGGGAATTc--->TGTTGGGAATTg
The light-inducible heterodimerizering proteins CRY2 and CIB1 are both from Arabidopsis thaliana.
pORE Expression Series Vector with BioBrick MCS (Kan reistance)
Vector for transformation of plants. Cloning in E. coli, transfer to Agrobacterium, integrates into plant genome. Kan resistance marker in bacteria, confers Kan resistance in plants. Has constitutive promoter upstream of multiple cloning site for easy expression of genes in plants. Conforms to Silver BioBrick Assembly Standard 21.
Modified vector pORE_O4 from the Arabidopsis Information Resource
Arabidopsis optimized Miraculin with StrepII C-terminus tag
Arabidopsis optimized Miraculin (BBa_K382021) with a StrepII tag for western blot analysis.
BBa_K382021 with biobricked strepII tag obtained from the Silver Laboratory.
Arabidopsis optimized Brazzein with a 2xYFP tag.
Brazzein was synthesized. YFP tag was added by cloning
PDK intron from pHANNIBAL vector for use in plant intron containing hairpin RNAi construction.
Arabidopsis optimized Miraculin with a 2xYFP tag.
Miraculin was synthesized. YFP tag was added by cloning
ihpRNA construct against Arabidopsis Ger3 allergen
Intron containing hairpin RNAi construct targeting Arabidopsis Germin 3 allergen sequence. Contains PDK intron from pHANNIBAL, assembled using Silver standard 21.
Eukaryotic Cryptochrome 2 (CRY2) ( a blue light stimulated photoreceptor)
Cryptochrome 2 (CRY2) is a blue light stimulated photoreceptor, when exposed to blue light, it would interact withthe N-terminal fragment of CIB1 (CIBN) . This part is full-length CRY2 CDS. Optogenetic systems enable precise spatial and temporal control of cell behavior. A light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light.This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive tCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes[1].
ihpRNA construct against Arabidopsis LTP Allergen
Intron containing hairpin RNA construct for RNAi knocktown of Lipid Transfer Protein allergen in Arabidopsis. Intron is PDK intron from pHANNIBAL assembled with 300bp of sense and antisense DNA from Arabidopsis genomic DNA assembled with Silver standard 21
Arabidopsis genomic DNA, pHANNIBAL vector
amiRNA Construct for carotene beta-ring hydroxylase (BETA-OHASE 1)
plant artificial microRNA construct for targeted knock-down of carotene beta-ring hydroxylase (BETA-OHASE 1). Needs to be transferred to a plant expression vector.
designed with WebMicroRNA designer http://wmd3.weigelworld.org/cgi-bin/webapp.cgi
modified from RS300, target sequences were synthesized as oligos,
pORE Open Series Vector with BioBrick MCS (Basta resistance)
Vector for transformation of plants. Cloning in E. coli, transfer to Agrobacterium, integrates into plant genome. Kan resistance marker in bacteria, confers Basta resistance in plants. Has no promoter or reporter.
Modified vector pORE_O1 from the Arabidopsis Information Resource
pORE Expression Series Vector with BioBrick MCS, Basta resistance
Vector for transformation of plants. Cloning in E. Coli, transfer to agrobacteria, integrates in the plant genome. Kan resistance in bacteria, confers Basta resistance in plants. Contains the pENTCUP promoter upstream of MCS for constitutive expression of gene of interest.
Modified from pORE_E3 found in Arabidopsis Information Resource
pORE GFP Reporter Series Vector with BioBrick MCS (Kan resistance)
Vector for transformation of plants. Cloning in E. coli, transfer to Agrobacterium, integrates into plant genome. Kan resistance marker in bacteria, confers Kan resistance in plants. Has constitutive plant promoter and in-frame GFP reporter. Conforms to Silver BIoBrick Assembly Standard 21.
Modified vector pORE_R2 from the Arabidopsis Information Resource
Intron containing hairpin RNAi construct targeting Arabidopsis Lipid Transfer Protein allergen. PDK intron from pHANNIBAL, 300 bp of LTP sense and antisense from Arabidopsis genomic DNA.
pORE Open Serices Vector with BioBrick MCS (Kan resistance)
Vector for transformation of plants. Cloning in E. coli, transfer to Agrobacterium, integrates into plant genome. Kan resistance marker in bacteria, confers Kan resistance in plants. Has no promoter or reporter. Conforms to Silver BioBrick Assembly Standard 21.
Miraculin is a 'flavor inverting' protein, found naturally in the fruit of the plant fruit of Synsepalum dulcificum. Not sweet by itself, miraculin binds to taste receptors on the tongue, possibly altering the structure of the receptors and causing traditionally 'sour' flavors to be received as 'sweet'.
Synthesized by Mr. Gene, optimized for expression in Arabidopsis. Derived from Genbank sequence BAH84844
Coding region for cellulose binding domain. When fused to a protein, protein attaches to cellulose
Domain derived from cellulose binding protein from Chlostridium cellulovorans(WP_013291799).
Arabidopsis optimized Brazzein with StrepII C-terminus
Arabidopsis optimized Miraculin with StrepII N-terminus tag
amiRNA Construct for lycopene epsilon cyclase (LUT2)
Artificial micro RNA targeting the Arabidopsis LUT2 gene.
Designed with the artificial Web MicroRNA Designer (http://wmd3.weigelworld.org/cgi-bin/webapp.cgi?page=Home;project=stdwmd)
Modified from RS300 to target the Arabidopsis LUT2.
Encodes the N-terminal fragment of transcription factor CIB1 (cryptochrome-interacting basic-helix-loop-helix). CIB1 interacts with CRY2 (cryptochrome 2) in a blue light-specific manner in yeast and Arabidopsis cells, and it acts together with additional CIB1-related proteins to promote CRY2-dependent floral initiation. CIB1 positively regulates FT expression. Cryptochrome 2 (CRY2) is a blue light stimulated photoreceptor, when exposed to blue light, it would interact withthe N-terminal fragment of CIB1 (CIBN) . Optogenetic systems enable precise spatial and temporal control of cell behavior. A light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light.This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive tCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes[1].
Top metal binding protein. Binds zinc, arsenic, copper and cadmium.
Multiple domain protein. Domains bind zinc, arsenic, copper and cadmium. Sequences derived from metallothioneins and transcription factors of different organisms. Arsenic, zinc and cadmium binding regions are full length metallothioneins. Zinc binding region derived from a transcritpion factor binding domain. Different protein binding regions are separated with glycerin-serine spacers to improve domain folding. Metal binding regions consists of disulfide bonds. For expression in E. coli disulfide bond forming strains are useful (e. g. Origami).
Arsenic (CAA06729), zinc (CAD88267) and cadmium (AAN06822) metallothioneins are full length. Copper AAA34542)domain is partial.
pENTCUP2 promoter for the constitutive expression of genes in plants.
Amplified from PCR from the pORE E3 plasmid.
Derived from the pORE vector series (see http://2010.igem.org/Team:Harvard).