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Open Enzymes

Open Enzymes

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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.



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

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Joel Graff jgraff {at} mtech {dot} edu United States
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Phil Roche philroche365 {at} gmail {dot} com Canada
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Marko Komlosh markokomlosh {at} gmail {dot} com Serbia
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Friede van der Berg 21555230 {at} sun {dot} ac {dot} za South Africa
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Maan Neamah maanm.neamah {at} uokufa {dot} edu {dot} iq Iraq
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Alex Nguyen alex.nguyenba {at} utoronto {dot} ca Canada
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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
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