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