The Open Enzyme Collection consists of 81 genes for enzymes that are "workhorses" of molecular biology, including DNA polymerases, RNA polymerases, DNA ligases, reverse transcriptases and restriction enzymes. The enzymes are useful in both basic and applied fields as diverse as genomics, diagnostics, biodiversity, synthetic biology, DNA origami, immunology, biochemistry and more.
The foundational technologies enabled by this collection of include polymerase chain reaction (PCR), cloning, isothermal amplification, reverse transcription of RNA into DNA, and dNTP synthesis. Additionally, the Open Enzyme collection contains enzyme genes for fixing damaged and degraded DNA prior to amplification, enzymes for cutting protein sequences to remove unwanted parts, and DNA-binding proteins that can be fused with other enzymes to improve functionality.
The Open Bioeconomy Lab designed and codon-optimized the Open Enzymes parts for expression in Escherichia coli bacteria. All enzymes are obtained from expired patents or for various reasons are not encumbered by patent rights.
What can the collection be used for?
Researchers can use the collection as the basis of manufacturing their own enzymes by cloning them into an expression vector of their choice and then transforming into E. coli bacteria. This can help reduce lab costs and overcome supply chain issues, particularly in areas of the world where enzymes are harder to obtain. The enzymes can also be used as a base to improve functionality or add new functions through genetic engineering and directed evolution.
Open Enzymes are not provided in expression vectors. In order to produce an enzyme, you must subclone your gene of interest into an expression vector. Open Enzymes are also compatible with the E. coli Protein Expression Toolkit (all parts have CE overhangs). More information about the collection can be obtained from the Open Bioeconomy Lab.
This product is made available under the unilateral OpenMTA.
Some or all of these items are for use only as permitted by a research exemption.
Instructions for Use:
Open Enzymes is Shipped as purified and dried down plasmid DNA stained with Cresol Red in one 96-well plate. Each well contains approximately 50 ng of DNA. Cresol Red will not impact plasmid transformation.
Transform into Top10 E. coli or equivalent. Pick a colony and culture at 37C overnight, with shaking at around 300rpm. The selection marker for all parts is ampicillin.
You may find an expression guide on the Open Bioeconomy Lab website.
The current KOD polymerase gene (Well F1) produces a nonfunctional protein. We are working to replace the polymerase gene with a functional version. Thank you to Tomas Moravec for pointing out the issue!
If you have already received the collection, check out the plate layout.
1 Plate in Set
This kit is included in kit plate 2 of the 2022 iGEM distribution. The plate map for kit plate 2 of the iGEM 2022 distribution can be found here.
Prior Kit Versions:
If your plate is labeled "Open Enzymes Part 1" or "openenzymes_1," you can find a plate map here.
If your plate is labeled "Open Enzymes Part 2," you can find a plate map here.
Genes
| Gene | Name | Freegenes ID |
|---|---|---|
| 9N7polA | 9°N-7 DNA polymerase | BBF10K_003247 |
| K12polLF | DNA Polymerase I, Large (Klenow) Fragment | BBF10K_003248 |
| K12polLF (exo) | DNA Polymerase I, Large (Klenow) Fragment (3′→5′ exo-) | BBF10K_003249 |
| ECOpolA | E. coli DNA Polymerase I | BBF10K_003250 |
| KTHEAQpol | KlenTaq1 polymerase | BBF10K_003251 |
| KODpol | KOD DNA polymerase | BBF10K_003252 |
| Pwopol | Pwo DNA Polymerase | BBF10K_003254 |
| GBDpol (exo) | Pyrococcus Sp. Heat-Stable (exo–) DNA Polymerase (Deep Vent™ (exo–) DNA Polymerase at NEB) | BBF10K_003255 |
| GBDpol | Pyrococcus Sp. Heat-Stable DNA Polymerase (Deep Vent™ at NEB) | BBF10K_003256 |
| THEAQpolA | Taq DNA Polymerase | BBF10K_003257 |
| Tlipol (exo) | Tli DNA polymerase (exo-) (available from NEB as VentR® (exo-) DNA Polymerase) | BBF10K_003259 |
| TthpolA | Tth DNA polymerase | BBF10K_003260 |
| Bstpol | Bst DNA Polymerase, Full Length | BBF10K_003261 |
| BstpolLF | Bst DNA Polymerase, Large Fragment | BBF10K_003262 |
| BsupolLF | Bsu DNA Polymerase I, Large Fragment | BBF10K_003263 |
| phi29pol | phi29 DNA Polymerase | BBF10K_003264 |
| 9N7polA (CT) | 9°N-7 DNA Polymerase chain terminating (NEB > Therminator DNA Pol) | BBF10K_003265 |
| dbh | Sulfolobus DNA Polymerase IV | BBF10K_003266 |
| T4gene43 | T4 DNA Polymerase | BBF10K_003267 |
| T7gene5 | T7 DNA Polymerase (unmodified) | BBF10K_003268 |
| T5gene122 | T5 DNA Polymerase | BBF10K_003269 |
| T4gene30 | T4-DNA Ligase | BBF10K_003270 |
| ECOligA | E. coli DNA Ligase | BBF10K_003271 |
| HSlig3 | DNA Lligase III | BBF10K_003272 |
| HSlig4 | DNA Ligase IV | BBF10K_003273 |
| 9N7lig | 9 Degrees North DNA Ligase | BBF10K_003274 |
| Pfulig | Pfu DNA Ligase | BBF10K_003275 |
| THEAQlig | Taq DNA Ligase | BBF10K_003276 |
| T4gene63 | T4 RNA Ligase 1 | BBF10K_003277 |
| ALPI | CIP (calf intestinal phosphatase) | BBF10K_003278 |
| RNAse A | RNAse A | BBF10K_003279 |
| RNAse H | RNAse H | BBF10K_003280 |
| EcoRIR | EcoRI restriction enzyme | BBF10K_003281 |
| EcoRIM | EcoRI methyltransferase | BBF10K_003282 |
| PstIR | PstI restriction enzyme | BBF10K_003283 |
| PstIM | PstI methyltransferase | BBF10K_003284 |
| MMLV_RT (mut H) | Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase (RNAse H deactivated by 3 mutations) | BBF10K_003285 |
| MMLV_RT (lack H) | Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase RNaseH - (lacking RNaseH domain) (SuperScriptII) | BBF10K_003286 |
| TthRT | Thermus thermophilus (Tth) RT | BBF10K_003287 |
| T3RNAp | T3 RNA Polymerase | BBF10K_003289 |
| T7RNAp | T7 RNA Polymerase | BBF10K_003290 |
| SP6RNAp | SP6 RNA Polymerase | BBF10K_003291 |
| Eco31I | Eco31I restriction enzyme | BBF10K_003292 |
| Eco31IA | Eco31IA methyltransferase | BBF10K_003293 |
| Eco31IB | Eco31IB methyltransferase | BBF10K_003294 |
| SapIR | SapI restriction enzyme | BBF10K_003295 |
| SapIM1 | M1.SapI methyltransferase | BBF10K_003296 |
| SapIM2 | M2.SapI methyltransferase | BBF10K_003297 |
| XbaIR | XbaI restriction enzyme | BBF10K_003298 |
| XbaIM | M.XbaI methyltransferase | BBF10K_003299 |
| NotIR (GC) | NotI restriction enzyme | BBF10K_003300 |
| NotIM | M.NotI methyltransferase | BBF10K_003301 |
| SfiIR (cDNA) | SfiI (used to create cDNA libraries) | BBF10K_003302 |
| SfiIM | M.SfiI methyltransferase | BBF10K_003303 |
| EcoRV | EcoRV restriction enzyme | BBF10K_003304 |
| EcoRVM | M.EcoRV methyltransferase | BBF10K_003305 |
| NcoIR | NcoI restriction enzyme | BBF10K_003306 |
| NcoIM | M.NcoI methyltransferase | BBF10K_003307 |
| HindIIIR | HindIII restriction enzyme | BBF10K_003308 |
| HindIIIM | M.HindIII methyltransferase | BBF10K_003309 |
| DpnIR | DpnI restriction enzyme | BBF10K_003310 |
| Bst HF | Bst polymerase, High fidelity | BBF10K_003311 |
| ETSSB | Extreme Thermostable Single-Stranded DNA Binding Protein | BBF10K_003312 |
| T5gene12/13 | T5 Exonuclease | BBF10K_003313 |
| TthligK294R | Tth DNA Ligase K294R | BBF10K_003314 |
| dromedNK | dNK from drosophila | BBF10K_003315 |
| TEV protease | TEV protease | BBF10K_003316 |
| T4 PNK | T4 Polynucleotide Kinase | BBF10K_003317 |
| T4-BGT | T4 Phage β-glucosyltransferase | BBF10K_003318 |
| T7EI | T7EI (T7 Endonuclease I) | BBF10K_003319 |
| T4 PGD | T4 PGD (T4 Endonuclease V) | BBF10K_003320 |
| T4 g32 | T4 gene 32 | BBF10K_003321 |
| P45 | P45 (ArchaeMaxx/PfuTurbo) | BBF10K_003322 |
| BovDNTT | Bovine DNTT | BBF10K_003323 |
| DNA Topoisomerase 1B Vaccinia Virus | DNA Topoisomerase 1B Vaccinia Virus | BBF10K_003324 |
| lambda red operon | lambda red operon (exo, bet, gam) | BBF10K_003325 |
| UDG | Uracil DNA glycosylase | BBF10K_003326 |
| Fpg | Formamidopyrimidine DNA Glycosylase | BBF10K_003327 |
| Endonuclease IV E. coli | Endonuclease IV E. coli | BBF10K_003328 |
| Endonuclease VIII E. coli | Endonuclease VIII E. coli | BBF10K_003329 |
| Exonuclease III E. coli | Exonuclease III E. coli | BBF10K_003330 |
Download all of this information as a CSV from our GitHub.
Bionet
The bionet enables open peer-peer exchange of functional biomaterials and associated data. This product may also be available from bionet nodes that are more convenient to you. Here are other bionet nodes who may be willing to provide you this specific product.| Name | Contact | Country |
|---|---|---|
| Tri Ngo | NA | United States |
| Stephen Klusza | smklusza {at} gmail {dot} com | United States |
| Codruta Ignea | codruta.ignea {at} mcgill {dot} ca | Canada |
| Aileen Button | acbutton {at} ucsd {dot} edu | United States |
| Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org | United States |
| Florian Leiß-Maier | florian.leiss-maier {at} tum {dot} de | Germany |
| Long Nguyen | Lnguyen2 {at} ufl {dot} edu | United States |
| Keith Gagnon | ktgagnon {at} gmail {dot} com | United States |
| Gülşen Günel | gulsen.gunel {at} ogr {dot} iu {dot} edu {dot} tr | Turkey |
| Jonathan Jonathan | Jonathancontact16 {at} gmail {dot} com | Indonesia |
| Benjamin Loang | benjamin.liang {at} uq {dot} edu {dot} au | Australia |
| Cihan Aydin | cihan.aydin {at} medeniyet {dot} edu {dot} tr | Turkey |
| Fernando Bustos | fernando.bustos {at} genosur {dot} com | Chile |
| Anil Challa | akchalla {at} uab {dot} edu | United States |
| Peter Rootes | root0059 {at} umn {dot} edu | United States |
| James Altamirano | James_altam {at} utexas {dot} edu | United States |
| Phil Jensik | NA | United States |
| Joey Riepsaame | joey.riepsaame {at} path {dot} ox {dot} ac {dot} uk | United Kingdom |
| N Sadananda Singh | nsslabresearch {at} gmail {dot} com | India |
| Nicholas White | nickk.white {at} gmail {dot} com | United States |
| Ellen Jorgensen | ejorgensen {at} biotechwithoutborders {dot} org | United States |
| Peer Schenk | reuben.brown {at} uqconnect {dot} edu {dot} au | Australia |
| Jorge Marchand | jmarcha {at} uw {dot} edu | United States |
| Maira Goytia | mgoytia {at} spelman {dot} edu | United States |
| Leandro Oliveira (NG) | ll637 {at} cam {dot} ac {dot} uk | United Kingdom |
| Yi Zhang | www.pharmozyme.com | United States |
| Arthur Zanetti Nunes Fernandes | fgueiros {at} iq {dot} usp {dot} br | United States |
| Vitaliy Strochkov | NA | Kazakhstan |
| Sreenivas Eadara | sreeni.eadara {at} gmail {dot} com | United States |
| Ivan Slovokhotov | ivan_slovohotov {at} mail {dot} ru | Russia |
| Alex Nguyen | alex.nguyenba {at} utoronto {dot} ca | Canada |
| Wei-Min Chang | WeiMinChang {at} tmu {dot} edu {dot} tw | Taiwan |
| Moritz Venne | moritz.venne {at} web {dot} de | Germany |
| Jacques Mathieu | mathieu {at} rice {dot} edu | United States |
| Simran Kushwaha | https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty | India |
| Wapouo Fadanka Stephane | mboalab {at} gmail {dot} com | Republic of Cameroon |
| Elizabeth Bilsland | bilsland {at} unicamp {dot} br | Brazil |
| Umair Hanif | umair.hanif {at} gdec {dot} edu {dot} pk | Pakistan |
| Friede van der Berg | 21555230 {at} sun {dot} ac {dot} za | South Africa |
| Muhammad Saad Rehmani | saadrehmani88 {at} hotmail {dot} com | China |
| Andrei Berasneu | a.berasneu {at} kleverlab {dot} pl | Poland |
| Tocvic Meng | Tocvic Meng \ Email: tocvic.meng {at} holocyte {dot} com \ website https://gmexpression {dot} com/ | Australia |
| Cauã Westman | caua.westmann {at} ieu {dot} uzh {dot} ch | Switzerland |
| Juan Sebastian Alvarez | juan.alvarez1 {at} ucalgary {dot} ca | Canada |
| elwi machado | elwi.machado {at} unisimonbolivar {dot} edu {dot} co | Colombia |
| Zhiyong Lei | zlei {at} umcutrecht {dot} nl | Netherlands |
| Dong Nguyen Tam | NA | Vietnam |
| Jianxiang Shi | jianxiangshi {at} zzu {dot} edu {dot} cn | China |
| Andrei Berasneu | andreyberesnevson {at} gmail {dot} com | Poland |
| Joao Vitor Dutra Molino | candidomolino {at} gmail {dot} com | United States |
| Nikolaus Dietz | nikolaus.dietz {at} unibas {dot} ch | Switzerland |
| Rodolfo Marcolino | rodolfo.marcolino {at} me {dot} com | Argentina |
| Esteban Erben | eerben {at} iib {dot} unsam {dot} edu {dot} ar | Argentina |
| Pratik Vyas | iampratikvyas {at} gamil {dot} com | India |
| Matthew Phanchana | matthew.pha {at} mahidol {dot} edu | Thailand |
| Amitabha Majumdar | Mamitava {at} nccs {dot} res {dot} in | India |
| Richard Jones | rpj1234rpj {at} gmail {dot} com | United States |
| Matias Cabeza | matiascabeza {at} gmail {dot} com | Argentina |
| Dayananda Chandrappa | nanda.daya {at} gmail {dot} com | India |
| Kefentse Arnold Tumedi | tumediak {at} yahoo {dot} com | Botswana |
| Ahmed Hegazy | a.hegazy {at} biotec {dot} rwth-aachen {dot} de | Germany |
| Ferdinand Molnár | NA | Kazakhstan |
| Jennifer Hsieh | j.hsieh {at} kurabiotech {dot} com | Chile |
| Hamidreza Karbalaeiheidari | karbalaei76 {at} gmail {dot} com | Canada |
| Stephen Klusza | StephenKlusza {at} clayton {dot} edu | United States |
| Alexander Iakovlev | atom.liquid {at} gmail {dot} com | Canada |
| Erman ÖZER | aracnoid {at} gmail {dot} com | Turkey |
| Ingrid Lorena Jaramillo | Lojadel2 {at} gmail {dot} com | Colombia |
| mehmet tardu | mtardu {at} gmail {dot} com | Turkey |
| Marko Komlosh | markokomlosh {at} gmail {dot} com | Serbia |
| Bunpote Siridechadilok | bunpote.sir {at} biotec {dot} or {dot} th | Thailand |
| Xingyue jiang | karma5781 {at} 21cn {dot} com | China |
| Watanyu Bunsermyos | watanyu.bu {at} ku {dot} th | Thailand |
| Alexandr Dorif | dorif11 {at} gmail {dot} com | Moldova, Republic of |
| Deepak Saini | deepaksaini {at} iisc {dot} ac {dot} in | India |
| Vincent Paris | NA | United States |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | Saudi Arabia |
| Paulina Ejsmont | research.lab {at} cri-paris {dot} org | France |
| Kulvinder Gill | kanwar {at} geneshifters {dot} com | United States |
| Tomasz Jurkowski | jurkowskit {at} cardiff {dot} ac {dot} uk | United Kingdom |
| Kenneth Frimpong | ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh | Ghana |
| Bronagh Hogan | wolfgang.fischle {at} kaust {dot} edu {dot} sa | Saudi Arabia |
| Romain Volmer | romain.volmer {at} envt {dot} fr | France |
| Jonathan Chevriau | j.chevriau {at} gmail {dot} com | Argentina |
| Tim Dobbs | tim.dobbs {at} cri-paris {dot} org | France |
| Emre Yörük | unalsergen {at} gmail {dot} com | Turkey |
| Manish Kushwaha | manish.kushwaha {at} inrae {dot} fr | France |
| Danielle Pedrolli | danielle.pedrolli {at} unesp {dot} br | Brazil |
| Shivang Joshi | https://www.bacto.bio | United Kingdom |
| Phil Roche | philroche365 {at} gmail {dot} com | Canada |
| Thooyavan George | www.hetrogenebiotech.com | India |
| Joel Graff | jgraff {at} mtech {dot} edu | United States |
| Oskar Zeballos | zebsamosk {at} gmail {dot} com | Bolivia |
| Prince Samoh | Samohprince.edem {at} gmail {dot} com | Ghana |
| Ben Thuronyi | bwt2 {at} williams {dot} edu | United States |
| Kheng Oon Low | khengoon {at} nibm {dot} my | Malaysia |
| Ben Davis | bed58 {at} aber {dot} ac {dot} uk | United Kingdom |
| Sarah Ware | sarah.ware {at} bioblaze {dot} org | United States |
| LUN CUI | luncui {at} cczu {dot} edu {dot} cn | China |
| Jose David Rosales | jdrr55 {at} yahoo {dot} com https://www {dot} linkedin {dot} com/in/david-rosales-574a7151/ | Venezuela |
| Maan Neamah | maanm.neamah {at} uokufa {dot} edu {dot} iq | Iraq |
| Juan Vicente Farizano | juan.farizano {at} fbqf {dot} unt {dot} edu {dot} ar | Argentina |
| Shobha KrupaRani | pshobha {at} ccmb {dot} res {dot} in | India |
| Kelly Gallagher | kelly.gallagher {at} oneonta {dot} edu | United States |
| Stijn Tobias de Vries | stijn.vries {at} mpi-marburg {dot} mpg {dot} de | Germany |
| Mariela Escobar | maru {at} michroma {dot} co | Argentina |
| Emin Bursa | NA | United States |
| Franklin Nobrega | f.nobrega {at} soton {dot} ac {dot} uk | United Kingdom |
| Julio Bonilla | jabonill {at} espol {dot} edu {dot} ec | Ecuador |
| Ashton Cropp | tacropp {at} vcu {dot} edu | United States |
| Fristot Elsa | elsa.fristot {at} cbs {dot} cnrs {dot} fr | France |
| Benjamin Arias | barias {at} alumni {dot} usfq {dot} edu {dot} ec | Ecuador |
| Han Teng Wong | wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg | Singapore |
| William Fried | uscigem {at} usc {dot} edu | United States |
| Jose Munoz | Jose.Munoz {at} northumbria {dot} ac {dot} uk | United Kingdom |
| Guillermo VegaLopez | guillermo.vega-lopez {at} fbqf {dot} unt {dot} edu {dot} ar | United States |
| Andrew Gray | andrewg {at} bioq {dot} org {dot} au | Australia |
| Shree Ram Singh | singhlabasu {at} gmail {dot} com | United States |
| Josephine Labos | N/A | Philippines |
| Matthew Thompson | matthew.thompson-12 {at} postgrad {dot} manchester {dot} ac {dot} uk | United Kingdom |
| Intan Taufik | i.taufik {at} sith {dot} itb {dot} ac {dot} id | Indonesia |
| Suresh Arakera | sbarakera {at} kud {dot} ac {dot} in | India |
| Jehovani Lopez | jlopez {at} genspace {dot} org | United States |
| Guillermo VegaLopez | guillermo.vega-lopez {at} fbqf {dot} unt {dot} edu {dot} ar | Argentina |
| Ian Cubit | Djcubit {at} nycap {dot} rr {dot} com | United States |
| David Truong | www.truonglab.com | United States |
| Jimmy Linhares | Synbioamazonas {at} gmail {dot} com | Brazil |
| Aleksandr Shilovich | mercurialbadger {at} yandex {dot} ru | Russia |
| semih cagan | semihcgn {at} gmail {dot} com | Turkey |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | United States |
| Sarah Turcotte | turcottes {at} cbsnews {dot} com | United States |
| Cole LaBonne | colelabonne {at} hotmail {dot} com | United States |
| Norhan Hassan | norhan.hassan {at} kaust {dot} edu {dot} sa | Saudi Arabia |
| Kristel Alman | kristel.alman {at} ut {dot} ee | Estonia |
| Chris Badenhorst | chris.badenhorst {at} uni-greifswald {dot} de | Germany |
| Marc Juul | marc {at} juul {dot} io | United States |
| Bingbing Wan | wanb {at} sjtu {dot} edu {dot} cn | China |
| Durga Burande | burandedurga22 {at} gmail {dot} com | India |
| Dayananda Chandrappa | e2ebiotech {at} gmail {dot} com | India |
| Muhammad Shoaib | mshoaib {at} lums {dot} edu {dot} pk | Pakistan |
| Majed Alghamdi | mafa2 {at} le {dot} ac {dot} uk | United Kingdom |
| Adrian Filips | tada {at} specyal {dot} com | Canada |
| Matthew Knuesel | mknuesel {at} hudsonalpha {dot} org | United States |
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