...Synthetic Biology: Assembly Standards Guide SynBio Resources SynBio Home Browse All SynBio Assembly Standards ...cloning strategies for the assembly of synthetic parts include Gibson assembly and type IIS systems such...Assembly standards define standard prefix and suffix sequences that make it easy to combine components...networks. This is accomplished through the use of assembly standards – combinations of restriction sites ...prefix and suffix sequence at either end of the assembly, even as more parts are added. Therefore, the ...The following table provides information for the assembly standards most commonly used by part registries...primer design. The restriction enzymes used for assembly are the prohibited sites and must be omitted from...

...subfamily C, member 3 NKG2-E, NKG2E KLRC4 killer cell lectin-like receptor subfamily C, member 4 FLJ17759...subfamily D, member 1 CD94 LGMN legumain AEP, LGMN1, PRSC1 LTA lymphotoxin alpha (TNF superfamily, member 1) LT...family A, member 2 PDA2, PDI, PDIP, PDIR PDIA3 protein disulfide isomerase family A, member 3 ER60, ERp57...FLJ12287, RP35, SEMAB, SEMB SEMA4B sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and short...MARVEL transmembrane domain containing 2 CKLFSF2, MGC39436 CMTM3 CKLF-like MARVEL transmembrane domain...CKLF-like MARVEL transmembrane domain containing 4 CKLFSF4 CMTM5 CKLF-like MARVEL transmembrane domain containing...MARVEL transmembrane domain containing 7 CKLFSF7, FLJ30992 CMTM8 CKLF-like MARVEL transmembrane domain...

... D, member 1 NR1D2 Plasmids NR1D2, RVR, BD73, EAR-1R nuclear receptor subfamily 1, group D, member 2 NR1H3...Browse the nuclear receptor plasmid collection assembled in collaboration with the Nuclear Receptor Signaling... domain by author. Source: Wikipedia, accessed November 6, 2013. Author: Boghog2 Nuclear receptors are...research labs from around the globe, Addgene has assembled a Nuclear Receptor plasmid collection for the ...NROB1, SRXY2 nuclear receptor subfamily 0, group B, member 1 NR1D1 Plasmids NR1D1, EAR1, hRev, THRA1, THRAL...LXRA, RLD-1 nuclear receptor subfamily 1, group H, member 3 NR2E1 Plasmids NR2E1, RP3-429G5.1, TLL, TLX, ..., XTLL nuclear receptor subfamily 2, group E, member 1 NR2F1 plasmids NR2F1, COUP-TFI, EAR-3, EAR3, ERBAL3...

...PI FusX TALEN Assembly System Contains 336 pFusX clones allowing for Golden Gate assembly of 15-16 RVD ...expression. Feng Zhang LIC TAL Effector Assembly Kit Assembly by ligation-independent cloning (LIC). Validated...plasmids of pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly. Kiran Musunuru...expression of Voytas Golden Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN...accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays. TALEN...Bogdanove Joung Lab TAL Effector Engineering Reagents Assembly via serial ligation. Validated in zebrafish somatic...Ekker Platinum Gate TALEN Kit Platinum TALENs assembled by Golden Gate. Validated in several species. ...

...Parkinson's Michael J Fox Foundation MJFF 40427 pEMB017 NP_9409803_LRRK2_1867_2161_L1914N_V2048T_F2083Y...Parkinson's Michael J Fox Foundation MJFF 40499 pEMB018 / LRRK2_BC_1300_2136 LRRK2 His polH Parkinson's...'s Michael J Fox Foundation MJFF 40500 pEMB018 / LRRK2_BC_1300_2148 LRRK2 His polH Parkinson's Michael...Michael J Fox Foundation MJFF 40501 pEMB018 / LRRK2_BC_1300_2170 LRRK2 His polH Parkinson's Michael J Fox Foundation... Foundation MJFF 40502 pEMB018 / LRRK2_BC_1300_2210 LRRK2 His polH Parkinson's Michael J Fox Foundation...Foundation MJFF 40503 pEMB018 / LRRK2_BC_1309_2136 LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40504... 40504 pEMB018 / LRRK2_BC_1309_2170 LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40505 pEMB018...

...37351 pQC membrane TdTomato IX Membrane Palmitoylation TdTomato Connie Cepko 22479 FUmGW Membrane Palmitoylation...pCAG-mGFP Membrane Palmitoylation sequence from GAP43 EGFP Connie Cepko 61099 Lck-GFP Membrane Palmitoylation...Mitochondria-Outer Membrane TOMM20 mCherry* Michael Davidson 118736 pBOB-CARMIL2 BH domain-GFP Plasma Membrane CARMIL2...protein of interest with known markers. Addgene has assembled a collection of fluorescent fusion proteins that... Michael Davidson 98822 Lck-mTurquoise2 Plasma Membrane Lck N-terminal lipidation signal mTurquoise2 Dorus... Dorus Gadella 98821 Lck-mScarlet-I Plasma Membrane Lck N-terminal lipidation signal mScarlet-I Dorus ... Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer Membrane Mff acGFP1 Gia Voeltz 158007 pCMV-mGold-Mito-N...

...And Ligation) assembly method (Sander et al., Nat Biotechnol. 2011), the REAL-Fast assembly method (Reyon...or REAL-Fast assembly process or lists of plasmid ID numbers required for FLASH assembly. The software... TAL effectors, including designed TALENs, and assembly methods for REAL, REAL-Fast and FLASH Genome...Ligation-based Automatable Solid-phase High-throughput) assembly method (Reyon and Tsai et al., Nat Biotechnol ...Biotechnol 2012). All three of these platforms assemble TAL effector repeat arrays on a single architectural framework...expression plasmids using REAL, REAL-Fast or FLASH assembly. This new version of ZiFiT Targeter enables users...these reagents are provided below: Joung Lab REAL Assembly TALEN Kit Individual REAL TALE Repeat Plasmids...

...separate vectors and then assembled into one construct using Golden Gate assembly. Gersbach MuLE System Mammalian...expression cassettes can be assembled using Golden Gate cloning or the Gibson Assembly. Chen Boosting CRISPR...this Golden Gate and Gateway based assembly method: plasmids for assembly of single gRNA vectors with various...Expression System Description PI Multiplex CRISPR/Cas9 Assembly System Kit Mammalian A system for constructing...-gRNA genes (PTGs) and then PTG cassettes are assembled into a Cas9 expressing vector. Yang A Robust CRISPR...multiple sgRNA expression cassettes, which can be assembled into the binary CRISPR/Cas9 vectors in one round...round of cloning by Golden Gate ligation or Gibson Assembly Liu A CRISPR/Cas9 toolbox for multiplexed plant...

...trafficking to the plasma membrane Acute disruption of the synaptic vesicle membrane protein synaptotagmin...measurements of membrane charges A FRET sensor enables quantitative measurements of membrane charges in live...Calcium Calcium sensor in astrocytes (GCaMP6f), membrane-tethered Khakh Lab pZac2.1 gfaABC1D-lck-GCaMP6f...2018 Sep;21(9):1272-1280. Lin Tian Calcium Plasma membrane targeted GCaMP6s calcium sensor A polarized Ca...+) Dynamics in the Mitochondrial Matrix and Intermembrane Space with the GZnP2 Sensor. ACS Chem Biol. ...sensor iATPSnFR1.0 and iATPSnFR1.1 (cytoplasmic, membrane-displayed, expression in neurons, astrocytes) ...reduced ph interference demonstrates augmented transmembrane chloride movement by gerbil prestin (SLC26a5...

...Finger Consortium Originally published in Addgene's December 2011 Newsletter Overview of TALEN Technology Imagine...can be made. Intuitively, one would expect that assembling relatively small, repeat-laden DNA regions into...from the University of Minnesota, arrays can be assembled in a matter of days. In an open access Nucleic...recently developed Golden Gate cloning method to assemble “multiple DNA fragments in an ordered fashion ...custom arrays consisting of 12-31 repeats can be assembled and inserted into a variety of backbones in just... in August, uses a serial ligation protocol to assemble arrays. Dr. Joung, co-founder of the Zinc Finger...early in 2011 and they described their kit for assembling TAL effector arrays in a 2012 issue of Nature...

... by REAL or REAL-Fast assembly or names of plasmids required for FLASH assembly. ID Plasmid Activation...expression vectors for TAL effector repeat arrays assembled using the REAL, REAL-Fast or FLASH methods. TALengineering...fragments encoding TAL effector repeat arrays (assembled using the REAL, REAL-Fast or FLASH methods) can...Additional details about the REAL, REAL-Fast, and FLASH assembly methods can be found on the TALengineering website...amino acid sequences (i.e. only the details of the assembly are different among the three methods, the final...

...pluripotency during embryonic development. This “reprogramming” of the cell to an embryonic state holds immense...cell lineages. Pluripotent stem cells, such as embryonic cells, can potentially give rise to any other ...limited to related cell lineages. Pluripotent embryonic and multipotent adult stem cells have been studied...removes the possible ethical concerns related to embryonic stem cell use and has the major advantage that...Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors...Mouse Direct Reprogramming of Fibroblasts into Embryonic Sertoli-like Cells by Defined Factors. Cell Stem...via microRNA-dependent and -independent pathways. EMBO Mol Med. 2017 Aug;9(8):1117-1131. Parmar Fibroblasts...

...Transfer proteins from the gel to a membrane. Incubate the membrane with an unconjugated primary antibody...block the membrane to remove excess antibody and prevent unwanted binding. Incubate the membrane with a ...Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane using either X-ray...tears, saliva, mucus) where it protects mucosal membranes Dimer with a valency of 4 IgA1; IgA2 IgD Coexpressed...short peptide linkers. Diabodies can be used for assembling protein nanostructures. Figure 2: An antibody...recognizes your primary antibody source species. Remember that recombinant antibodies will also have a source...

...regulators of membrane traffic, they also play critical roles at cilia, centrosomes, the plasma membrane, lipid... collection of ARF family plasmids. ARF family members include regulatory GTPases, GEFs, and GAPs. Plasmid...regulatory GTPases have long been characterized as members of the RAS superfamily, though recently have been...signaling activity. Locations of ARF Superfamily members. Figure from Szutl, et al 2019 PMID: 31084567 .... . More information about ARF Superfamily members can be found in: ARF GTPases and their GEFs and GAPs...

...e.g. maize and Arabidopsis) and vectors for easy assembly of one or more gRNA expression cassettes - Qi-Jun...intronized Cas9 series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs - Sylvestre Marillonnet...fluorescent seed selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants...creating plant expression vectors: A modular plasmid assembly kit for multigene expression, gene silencing, ...Parts II and Infrastructure Kit for hierarchical assembly of plant multigene constructs - Johannes Stuttmann...cloning chassis and set of functional parts for assembling plant transformation vectors - Daniel Voytas....

...expression vectors for TAL effector repeat arrays assembled using the REAL, REAL-Fast or FLASH methods...fragments encoding TAL effector repeat arrays (assembled using the REAL, REAL-Fast or FLASH methods) can...Additional details about the REAL, REAL-Fast, and FLASH assembly methods can be found on the TALengineering website...amino acid sequences (i.e. only the details of the assembly are different among the three methods, the final...REAL TALE Repeat Plasmids in the Joung Lab REAL Assembly TALEN Kit . Vectors encoding wild-type FokI nuclease...

...Adenoviral A block for the AdenoBuilder genome assembly system. Block 6 with deletion in E3 and insertion...Adenoviral A block for the AdenoBuilder genome assembly system; consolidates the regions present in pAd5...Adenoviral A block for the AdenoBuilder genome assembly system. Block 1 with deletion in E1 and insertion...adenovirus serotype 5 (HAdV5) genome that can be assembled in a one-tube reaction. ID Kit Description PI ... Plasmids contain modular inserts for seamless assembly of multiple mutations, alterations, or transgene...

...deposited tools and resources, or read our guide on assembly standards. Browse Plasmids by Microbe System Plasmids...chromosomes - Endy and Shearwin Labs Ralstonia-GG-Kit : Assembly of Ralstonia solanacearum species complex (Rssc...lipolytica Golden Gate tool kit : Integrative vector assembly of up to three transcription units in the oleaginous...circuit design in E. coli - Freemont Lab MoClo-YTK : Assembly of single and multi-gene constructs for S. cerevisiae...perform your synbio experiment. Synthetic Biology Assembly Standards Guide Bacterial Expression Systems :...

...Methods, September 2007, Vol. 4 No. 9, pp. 741-6 Shemiakina et al. : Nature Communications, November 2012,...Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you...result in a diverse set of colors. Addgene has assembled a collection of empty plasmid backbones with different...Stepanenko et al. : BioTechniques, Vol. 51, No. 5, November 2011, pp. 313–327 Scholarpedia : Entry by Robert...Shaner et al. : Journal of Cell Science, Vol. 120, December 2007, pp. 4247-4260 Shaner et al. : Nature Methods...

...method Modular Assembly Engineering Reagents - Engineer zinc finger arrays via Modular Assembly method Nuclease...finger proteins deposited by Zinc Finger Consortium members like the Joung Lab, Voytas Lab, and Wolfe Lab Plasmid... engineered zinc finger technology. Consortium members Keith Joung and Daniel Voytas have deposited at...