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Species: Other
Genetic Insert: T4 PNK
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: The most commonly used polynucleotide kinase. Used for: 5' phosphorylation of DNA/RNA for subsequent ligation; end labeling DNA or RNA for probes and DNA sequencing; removal of 3' phosphoryl groups.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165520 Copy
Species: Other
Genetic Insert: Tli DNA polymerase (exo-)
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: DNA polymerase; lacks 3' to 5' exonuclease activity. Intein encoded endonucleases are thought to mediate intein mobility by site-specific recombination initiated by endonuclease cleavage at the 'homing site' in gene that lack the intein. Comparable to VentR (exo-) DNA Polymerase at NEB.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165523 Copy
Species: Other
Genetic Insert: lambda red operon
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Operon containing Exo, Bet, and Gam. To use the lambda red recombineering system to modify your target DNA (including E. coli genome), electroporate a linear donor DNA substrate (either dsDNA or ssDNA) into E. coli expressing the lambda red enzymes. These enzymes then catalyze the homologous recombination of the substrate with the target DNA sequence (mutagenesis occurs in vivo). ~50 nucleotides of homology are required to the target site for recombination.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165521 Copy
Species: Other
Genetic Insert: Eco31IB
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Site-specific DNA methyltransferase 2; protects DNA from cleavage by Eco31I.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165527 Copy
Species: Other
Genetic Insert: SfiI (used to create cDNA libraries)
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Type II restriction enzyme that recognizes the double-stranded sequence GGCCNNNNNGGCC and cleaves after N-4.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165528 Copy
Species: Other
Genetic Insert: Pfu DNA Ligase
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: DNA ligase that seals nicks in double-stranded DNA during DNA replication, DNA recombination and DNA repair. Catalyzes template-dependent ligation at temperatures of about 30-80 degrees C, and substantially retains its catalytic activity when subjected to temperatures of about 85 degrees C to about 100 degrees C.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165525 Copy
Species: Other
Genetic Insert: Taq DNA Polymerase
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: DNA polymerase used in (routine) PCR method because of its high thermostability, standard testing, screening and output testing. Has a relatively high error rate because it lacks 3'-5' exonuclease activity and proofreading functionality. 5'-3' flap endonuclease degrades displaced strand.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification.
Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/ and https://freegenes.github.io/genes/BBF10K_003257.html.
Proper citation: RRID:Addgene_165512 Copy
Species: Other
Genetic Insert: DNA Polymerase I, Large (Klenow) Fragment
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: DNA polymerase fragment that retains 5'-3' polymerase activity and 3'-5' exonuclease activity for removal of precoding nucleotides and proofreading, but loses 5'-3' exonuclease activity. Usable for synthesis of double-stranded DNA from single-stranded templates, filling in of receded 3' ends of DNA fragments to make 5' overhang blunt, digesting away protruding 3' overhangs, preparation of radioactive DNA probes.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165511 Copy
Species: Other
Genetic Insert: M.EcoRI
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Methylase that recognizes the double-stranded sequence GAATTC, causes specific methylation on A-3 on both strands, and protects the DNA from cleavage by the EcoRI endonuclease.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165515 Copy
Species: Other
Genetic Insert: M.NotI
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Site-specific DNA methyltransferase; protects DNA from cleavage by NotI.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165518 Copy
Species: Other
Genetic Insert: Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase (RNAse H deactivated by 3 mutations)
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Single 75 kDa monomer, cDNA synthesis; high enzyme activity and processivity.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165546 Copy
Species: Other
Genetic Insert: Extreme Thermostable Single-Stranded DNA Binding Protein
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: ssDNA binding protein, 9kDa. Features: Improves the processivity of DNA polymerase; involved in stabilization and marking of ssDNA structure; increases the yield and specificity of PCR; increases the yield and processivity of RT during RT-PCR; improves DNA sequencing through regions with strong secondary structure.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165549 Copy
Species: Other
Genetic Insert: EcoRV
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Type II restriction enzyme that recognizes the double-stranded sequence GATATC and cleaves after T-1.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165548 Copy
Species: Other
Genetic Insert: UDG
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Used in PreCR mix from NEB, Uracil DNA glycosylase; Monofunctional DNA glycosylase that catalyzes the hydrolysis of the N-glycosidic bond from deoxyuridine to release uracil; Active on ss and dsDNA; Heat inactivation not possible.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165531 Copy
Species: Other
Genetic Insert: T7 DNA Polymerase (unmodified)
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: DNA Polymerase that catalyzes the replication of T7 phage DNA during infection. The protein dimer has two catalytic activities: DNA polymerase activity and strong 3'-5' exonuclease. The high fidelity and rapid extension rate of the enzyme make it particularly useful in copying long stretches of DNA template.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165534 Copy
Species: Other
Genetic Insert: M.SfiI
Vector Backbone Description: Vector Backbone:pBuild; Vector Types:Synthetic Biology; Bacterial Resistance:Ampicillin
Comments: Site-specific DNA methyltransferase; protects DNA from cleavage by SfiI.
The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information can be found at https://www.addgene.org/depositor-collections/open-enzyme/.
Proper citation: RRID:Addgene_165538 Copy
Species: Other
Genetic Insert: HCoV-OC43 S (spike)
Vector Backbone Description: Vector Backbone:pDest-303; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32504802
Comments: DNA sequence optimized for mammalian expression
Proper citation: RRID:Addgene_166015 Copy
Species: Other
Genetic Insert: MERS-CoV S (spike)
Vector Backbone Description: Vector Backbone:pDest-303; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32504802
Comments: DNA sequence optimized for mammalian expression
Proper citation: RRID:Addgene_166013 Copy
Species: Other
Genetic Insert: SARS-CoV S (spike)
Vector Backbone Description: Vector Backbone:pDest-303; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32504802
Comments: DNA sequence optimized for mammalian expression
Proper citation: RRID:Addgene_166012 Copy
Species: Other
Genetic Insert: SARS-CoV-2 RBD
Vector Backbone Description: Vector Backbone:pDest-303; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:33248226
Comments: DNA sequence optimized for mammalian expression
Proper citation: RRID:Addgene_166019 Copy
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