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 |
|---|---|---|
| Joel Graff | jgraff {at} mtech {dot} edu | United States |
| Durga Burande | burandedurga22 {at} gmail {dot} com | India |
| Phil Roche | philroche365 {at} gmail {dot} com | Canada |
| Haoyong WANG | research | China |
| Franklin Nobrega | f.nobrega {at} soton {dot} ac {dot} uk | United Kingdom |
| Shivang Joshi | https://www.bacto.bio | United Kingdom |
| Ivan Slovokhotov | ivan_slovohotov {at} mail {dot} ru | Russia |
| Suresh Arakera | sbarakera {at} kud {dot} ac {dot} in | India |
| Tri Ngo | NA | United States |
| Mariela Escobar | maru {at} michroma {dot} co | Argentina |
| Jordan Gonzalez | jgonzalez {at} thecitizensciencelab {dot} org | United States |
| Nikolaus Dietz | nikolaus.dietz {at} unibas {dot} ch | Switzerland |
| Jimmy Linhares | Synbioamazonas {at} gmail {dot} com | Brazil |
| Jonathan LeCureux | jlecureu {at} svsuedu | United States |
| elwi machado | elwi.machado {at} unisimonbolivar {dot} edu {dot} co | Colombia |
| Muhammad Saad Rehmani | saadrehmani88 {at} hotmail {dot} com | China |
| Guillermo VegaLopez | guillermo.vega-lopez {at} fbqf {dot} unt {dot} edu {dot} ar | United States |
| Majed Alghamdi | mafa2 {at} le {dot} ac {dot} uk | United Kingdom |
| Moritz Venne | moritz.venne {at} web {dot} de | Germany |
| Florian Leiß-Maier | florian.leiss-maier {at} tum {dot} de | Germany |
| Wei-Min Chang | WeiMinChang {at} tmu {dot} edu {dot} tw | Taiwan |
| Vincent Paris | NA | United States |
| Amitabha Majumdar | Mamitava {at} nccs {dot} res {dot} in | India |
| Meng Lu | menglu {at} iastate {dot} edu | United States |
| Matias Cabeza | matiascabeza {at} gmail {dot} com | Argentina |
| Bronagh Hogan | wolfgang.fischle {at} kaust {dot} edu {dot} sa | Saudi Arabia |
| Jonathan Chevriau | j.chevriau {at} gmail {dot} com | Argentina |
| Ben Thuronyi | bwt2 {at} williams {dot} edu | United States |
| Tomasz Jurkowski | jurkowskit {at} cardiff {dot} ac {dot} uk | United Kingdom |
| Paulina Ejsmont | research.lab {at} cri-paris {dot} org | France |
| Erman ÖZER | aracnoid {at} gmail {dot} com | Turkey |
| N Sadananda Singh | nsslabresearch {at} gmail {dot} com | India |
| Intan Taufik | i.taufik {at} sith {dot} itb {dot} ac {dot} id | Indonesia |
| Peer Schenk | reuben.brown {at} uqconnect {dot} edu {dot} au | Australia |
| Tim Dobbs | tim.dobbs {at} cri-paris {dot} org | France |
| Dong Nguyen Tam | NA | Vietnam |
| Sreenivas Eadara | sreeni.eadara {at} gmail {dot} com | United States |
| Esteban Erben | eerben {at} iib {dot} unsam {dot} edu {dot} ar | Argentina |
| Jianxiang Shi | jianxiangshi {at} zzu {dot} edu {dot} cn | China |
| Ellen Jorgensen | ejorgensen {at} biotechwithoutborders {dot} org | United States |
| Wapouo Fadanka Stephane | mboalab {at} gmail {dot} com | Republic of Cameroon |
| Peter Rootes | root0059 {at} umn {dot} edu | United States |
| Bunpote Siridechadilok | bunpote.sir {at} biotec {dot} or {dot} th | Thailand |
| Deepak Saini | deepaksaini {at} iisc {dot} ac {dot} in | India |
| Ian Cubit | Djcubit {at} nycap {dot} rr {dot} com | United States |
| Adrian Filips | tada {at} specyal {dot} com | Canada |
| Shawn Lyons | smlyons1 {at} bu {dot} edu | United States |
| Joao Vitor Dutra Molino | candidomolino {at} gmail {dot} com | United States |
| Ahmed Hegazy | a.hegazy {at} biotec {dot} rwth-aachen {dot} de | Germany |
| Emre Yörük | unalsergen {at} gmail {dot} com | Turkey |
| Emin Bursa | NA | United States |
| Julio Bonilla | jabonill {at} espol {dot} edu {dot} ec | Ecuador |
| Jonathan Jonathan | Jonathancontact16 {at} gmail {dot} com | Indonesia |
| Watanyu Bunsermyos | watanyu.bu {at} ku {dot} th | Thailand |
| Shree Ram Singh | singhlabasu {at} gmail {dot} com | United States |
| Jose David Rosales | jdrr55 {at} yahoo {dot} com https://www {dot} linkedin {dot} com/in/david-rosales-574a7151/ | Venezuela |
| Andrei Berasneu | andreyberesnevson {at} gmail {dot} com | Poland |
| Romain Volmer | romain.volmer {at} envt {dot} fr | France |
| Ben Davis | bed58 {at} aber {dot} ac {dot} uk | United Kingdom |
| Nicholas White | nickk.white {at} gmail {dot} com | United States |
| Jorge Marchand | jmarcha {at} uw {dot} edu | United States |
| Bingbing Wan | wanb {at} sjtu {dot} edu {dot} cn | China |
| Marc Juul | marc {at} juul {dot} io | United States |
| Michael Powers | mpowers9304 {at} gmail {dot} com | United States |
| Tocvic Meng | Tocvic Meng \ Email: tocvic.meng {at} holocyte {dot} com \ website https://gmexpression {dot} com/ | Australia |
| James Altamirano | James_altam {at} utexas {dot} edu | United States |
| Jacques Mathieu | mathieu {at} rice {dot} edu | United States |
| Stephen Klusza | smklusza {at} gmail {dot} com | United States |
| Long Nguyen | Lnguyen2 {at} ufl {dot} edu | United States |
| Yi Zhang | www.pharmozyme.com | United States |
| Jehovani Lopez | jlopez {at} genspace {dot} org | United States |
| Simran Kushwaha | https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty | India |
| Shobha KrupaRani | pshobha {at} ccmb {dot} res {dot} in | India |
| Han Teng Wong | wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg | Singapore |
| Oskar Zeballos | zebsamosk {at} gmail {dot} com | Bolivia |
| Fernando Bustos | fernando.bustos {at} genosur {dot} com | Chile |
| Alexandr Dorif | dorif11 {at} gmail {dot} com | Moldova, Republic of |
| Prince Samoh | Samohprince.edem {at} gmail {dot} com | Ghana |
| Joey Riepsaame | joey.riepsaame {at} path {dot} ox {dot} ac {dot} uk | United Kingdom |
| Manish Kushwaha | manish.kushwaha {at} inrae {dot} fr | France |
| Ferdinand Molnár | NA | Kazakhstan |
| Andrew Gray | andrewg {at} bioq {dot} org {dot} au | Australia |
| Danielle Pedrolli | danielle.pedrolli {at} unesp {dot} br | Brazil |
| Cihan Aydin | cihan.aydin {at} medeniyet {dot} edu {dot} tr | Turkey |
| Benjamin Arias | barias {at} alumni {dot} usfq {dot} edu {dot} ec | Ecuador |
| Juan Vicente Farizano | juan.farizano {at} fbqf {dot} unt {dot} edu {dot} ar | Argentina |
| Aleksandr Shilovich | mercurialbadger {at} yandex {dot} ru | Russia |
| Benjamin Loang | benjamin.liang {at} uq {dot} edu {dot} au | Australia |
| Xingyue jiang | karma5781 {at} 21cn {dot} com | China |
| Andrei Berasneu | a.berasneu {at} kleverlab {dot} pl | Poland |
| Dayananda Chandrappa | e2ebiotech {at} gmail {dot} com | India |
| LUN CUI | luncui {at} cczu {dot} edu {dot} cn | China |
| Leandro Oliveira (NG) | ll637 {at} cam {dot} ac {dot} uk | United Kingdom |
| David Truong | www.truonglab.com | United States |
| Alexander Iakovlev | atom.liquid {at} gmail {dot} com | Canada |
| Keith Gagnon | ktgagnon {at} gmail {dot} com | United States |
| Norhan Hassan | norhan.hassan {at} kaust {dot} edu {dot} sa | Saudi Arabia |
| Chris Badenhorst | chris.badenhorst {at} uni-greifswald {dot} de | Germany |
| Thooyavan George | www.hetrogenebiotech.com | India |
| Maira Goytia | mgoytia {at} spelman {dot} edu | United States |
| Phil Jensik | NA | United States |
| Codruta Ignea | codruta.ignea {at} mcgill {dot} ca | Canada |
| Umair Hanif | umair.hanif {at} gdec {dot} edu {dot} pk | Pakistan |
| Ingrid Lorena Jaramillo | Lojadel2 {at} gmail {dot} com | Colombia |
| Matthew Knuesel | mknuesel {at} hudsonalpha {dot} org | United States |
| Kulvinder Gill | kanwar {at} geneshifters {dot} com | United States |
| Kelly Gallagher | kelly.gallagher {at} oneonta {dot} edu | United States |
| Sarah Ware | sarah.ware {at} bioblaze {dot} org | United States |
| Anil Challa | akchalla {at} uab {dot} edu | United States |
| Pratik Vyas | iampratikvyas {at} gamil {dot} com | India |
| Kefentse Arnold Tumedi | tumediak {at} yahoo {dot} com | Botswana |
| Jose Munoz | Jose.Munoz {at} northumbria {dot} ac {dot} uk | United Kingdom |
| Arthur Zanetti Nunes Fernandes | fgueiros {at} iq {dot} usp {dot} br | United States |
| Tal Globus | bio {at} talglobus {dot} com | United States |
| Matthew Phanchana | matthew.pha {at} mahidol {dot} edu | Thailand |
| Elizabeth Bilsland | bilsland {at} unicamp {dot} br | Brazil |
| Rodolfo Marcolino | rodolfo.marcolino {at} me {dot} com | Argentina |
| Fristot Elsa | elsa.fristot {at} cbs {dot} cnrs {dot} fr | France |
| Kenneth Frimpong | ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh | Ghana |
| Aileen Button | acbutton {at} ucsd {dot} edu | United States |
| Hamidreza Karbalaeiheidari | karbalaei76 {at} gmail {dot} com | Canada |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | Saudi Arabia |
| Josephine Labos | N/A | Philippines |
| Cauã Westman | caua.westmann {at} ieu {dot} uzh {dot} ch | Switzerland |
| Marko Komlosh | markokomlosh {at} gmail {dot} com | Serbia |
| Matthew Thompson | matthew.thompson-12 {at} postgrad {dot} manchester {dot} ac {dot} uk | United Kingdom |
| Friede van der Berg | 21555230 {at} sun {dot} ac {dot} za | South Africa |
| Cole LaBonne | colelabonne {at} hotmail {dot} com | United States |
| Maan Neamah | maanm.neamah {at} uokufa {dot} edu {dot} iq | Iraq |
| Jennifer Hsieh | j.hsieh {at} kurabiotech {dot} com | Chile |
| Muhammad Shoaib | mshoaib {at} lums {dot} edu {dot} pk | Pakistan |
| Ashton Cropp | tacropp {at} vcu {dot} edu | United States |
| William Fried | uscigem {at} usc {dot} edu | United States |
| Zhiyong Lei | zlei {at} umcutrecht {dot} nl | Netherlands |
| Vitaliy Strochkov | NA | Kazakhstan |
| Muzaffar Muminov | muminov.imbio {at} gmail {dot} com | Uzbekistan |
| Stephen Klusza | StephenKlusza {at} clayton {dot} edu | United States |
| Guillermo VegaLopez | guillermo.vega-lopez {at} fbqf {dot} unt {dot} edu {dot} ar | Argentina |
| semih cagan | semihcgn {at} gmail {dot} com | Turkey |
| Alex Nguyen | alex.nguyenba {at} utoronto {dot} ca | Canada |
| Juan Sebastian Alvarez | juan.alvarez1 {at} ucalgary {dot} ca | Canada |
| Gülşen Günel | gulsen.gunel {at} ogr {dot} iu {dot} edu {dot} tr | Turkey |
| mehmet tardu | mtardu {at} gmail {dot} com | Turkey |
| Stijn Tobias de Vries | stijn.vries {at} mpi-marburg {dot} mpg {dot} de | Germany |
| Kristel Alman | kristel.alman {at} ut {dot} ee | Estonia |
| Richard Jones | rpj1234rpj {at} gmail {dot} com | United States |
| Simon Hofer | soyouwantalab {at} gmail {dot} com | Austria |
| Kheng Oon Low | khengoon {at} nibm {dot} my | Malaysia |
| Ahmed Atef | ahmed_atefaig2 {at} yahoo {dot} com | United States |
| Sarah Turcotte | turcottes {at} cbsnews {dot} com | United States |
| Dayananda Chandrappa | nanda.daya {at} gmail {dot} com | India |
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