E. coli Protein Expression Toolkit

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Are you looking for an all-in-one toolkit to build up your dream construct to try expressing your enzymes of choice, tailored to your lab’s resources and needs? A toolkit containing different promoters, RBSs, linkers, tags, reporters? This collection might be for you.

A collection of >100 DNA parts designed to be assembled in one-pot reaction using BsaI-based Golden-Gate assembly. DNA parts are MoClo, SEVA, Loop and CIDAR assembly compatible. The collection includes:

3 promoters
7 RBS and insulators
9 purification tags, 3 localization signals
5 cleavage sites, flexible linkers
23 genes of which 5 are reporters and 18 are used in diagnostics
3 destination vectors

The Protein Expression Toolkit is available as purified nucleic acids.

What can it be used for:

Use the DNA parts of the collection to assemble the expression plasmids for the protein of your choice, tailored to the resources present in your lab. The collection enables thousands of combinations for each gene of choice. Full power of the collection can be explored using pipetting robots. The collection is compatible with all the CDSs present in the Open Enzyme collections.

Assembly Scheme

If you've already received the kit, check out the layout for Plate 1 and Plate 2.

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.

If you are interested in requesting additional parts to be added to this toolkit, complete this request form.

Genes

Gene	Name	NCBI ID	Freegenes ID
CE_meffBlue/Rtms5/ NF pocilloporin /Biobrick K1033902	CE_meffBlue/Rtms5/ NF pocilloporin /Biobrick K1033902		BBF10K_003332
CE_sarcGFP	CE_sarcGFP		BBF10K_003356
CE_beta-galactosidase	CE_beta-galactosidase		BBF10K_003366
AF_lacZ pDest	AF_lacZ pDest	N/A	BBF10K_003373
AE_lacZ pDest	AE_lacZ pDest	N/A	BBF10K_003375
AF_high copy (pUC) ori/KanR	AF_high copy (pUC) ori/KanR	N/A	BBF10K_003376
AF_medium copy (pET) ori/KanR	AF_medium copy (pET) ori/KanR	N/A	BBF10K_003377
AB_T7	AB_T7	N/A	BBF10K_003378
AB_T7_lacO	AB_T7_lacO	N/A	BBF10K_003379
AB_pTac	AB_pTac	N/A	BBF10K_003380
BT1_BCD2	BT1_BCD2	N/A	BBF10K_003384
BT1_Riboj54_F1	BT1_Riboj54_F1	N/A	BBF10K_003385
BT1_Riboj_B34	BT1_Riboj_B34	N/A	BBF10K_003386
BT1_BCD12	BT1_BCD12	N/A	BBF10K_003387
BT1_BCD23	BT1_BCD23	N/A	BBF10K_003388
BT1_BCD2_His	BT1_BCD2_His	N/A	BBF10K_003389
BT1_RiboJ_B34_His	BT1_RiboJ_B34_His	N/A	BBF10K_003390
BT1_BCD2_DsbA	BT1_BCD2_DsbA	N/A	BBF10K_003391
BT1_BCD2_OmpT	BT1_BCD2_OmpT	N/A	BBF10K_003392
BT1_BCD2_pelB	BT1_BCD2_pelB	N/A	BBF10K_003393
T1T2_Calm	T1T2_Calm	N/A	BBF10K_003395
T1T2_CBD	T1T2_CBD	N/A	BBF10K_003396
T1T2_Cex	T1T2_Cex	N/A	BBF10K_003397
T1T2_MBP	T1T2_MBP	N/A	BBF10K_003398
T1T2_R5	T1T2_R5	N/A	BBF10K_003399
T1T2_GST	T1T2_GST	N/A	BBF10K_003400
T1T2_CipA	T1T2_CipA	N/A	BBF10K_003401
T1T2_S-Tag	T1T2_S-Tag	N/A	BBF10K_003402
T1T2_GB1	T1T2_GB1	N/A	BBF10K_003403
T1T2_GS flexible	T1T2_GS flexible	N/A	BBF10K_003404
T1T3_GS flexible	T1T3_GS flexible	N/A	BBF10K_003405
T1C_GS flexible	T1C_GS flexible	N/A	BBF10K_003406
T1C_His_TEVcut	T1C_His_TEVcut	N/A	BBF10K_003407
BT2_BCD2_His	BT2_BCD2_His	N/A	BBF10K_003408
BT2_RiboJ_B34	BT2_RiboJ_B34	N/A	BBF10K_003409
BC_B0034	BC_B0034	N/A	BBF10K_003410
BC_B0032	BC_B0032	N/A	BBF10K_003411
BC_RiboJ_RBS64	BC_RiboJ_RBS64	N/A	BBF10K_003412
BC_RiboJ_B34	BC_RiboJ_B34	N/A	BBF10K_003413
BC_BCD23	BC_BCD23	N/A	BBF10K_003414
BC_BCD12	BC_BCD12	N/A	BBF10K_003415
BC_BCD2	BC_BCD2	N/A	BBF10K_003416
T2C_GS flexible	T2C_GS flexible	N/A	BBF10K_003417
T2C_TEVcut	T2C_TEVcut	N/A	BBF10K_003418
BC_BCD2_His_TEVcut	BC_BCD2_His_TEVcut	N/A	BBF10K_003419
BC_BCD2_His_fuGFP_TEVcut	BC_BCD2_His_fuGFP_TEVcut	N/A	BBF10K_003420
T2T3_int_∆I	T2T3_int_∆I		BBF10K_003421
T2T3_SUMO	T2T3_SUMO		BBF10K_003422
T2T3_fuGFP	T2T3_fuGFP		BBF10K_003423
T2T3_AmilCP	T2T3_AmilCP		BBF10K_003424
T2T3_GS flexible	T2T3_GS flexible		BBF10K_003425
T3C_SUMO	T3C_SUMO		BBF10K_003426
T3C_TEV	T3C_TEV		BBF10K_003427
T3C_int_∆I	T3C_int_∆I		BBF10K_003428
T3C_fuGFP	T3C_fuGFP		BBF10K_003429
T3C_GS flexible linker	T3C_GS flexible linker	N/A	BBF10K_003430
CD_OMMLV	CD_OMMLV		BBF10K_003431
CD_HIVRT	CD_HIVRT		BBF10K_003432
CD_Bst-LF	CD_Bst-LF		BBF10K_003433
CD_OpenVent	CD_OpenVent		BBF10K_003434
CD_RNAseInhibitor	CD_RNAseInhibitor		BBF10K_003436
CD_gp32	CD_gp32		BBF10K_003437
CD_BsuLF	CD_BsuLF		BBF10K_003438
CD_UvsY	CD_UvsY		BBF10K_003439
CD_UvsX	CD_UvsX		BBF10K_003440
CE_T7RNApol_Stop	CE_T7RNApol_Stop		BBF10K_003441
CD_PBCV-1_ligase	CD_PBCV-1_ligase		BBF10K_003442
CD_HRP	CD_HRP		BBF10K_003443
CD_TEV_protease	CD_TEV_protease		BBF10K_003444
CD_SUMO_protease	CD_SUMO_protease		BBF10K_003445
CD_Calmodulin	CD_Calmodulin		BBF10K_003446
CD_fuGFP	CD_fuGFP		BBF10K_003447
CD_AmilCP	CD_AmilCP		BBF10K_003448
CD_TthRNAseH	CD_TthRNAseH		BBF10K_003449
CD_EcRNAseH	CD_EcRNAseH		BBF10K_003450
DT4_Int_SaltS	DT4_Int_SaltS	N/A	BBF10K_003451
DT4_Int_ThiolS	DT4_Int_ThiolS	N/A	BBF10K_003452
DT4_TEV	DT4_TEV	N/A	BBF10K_003453
DT4_fuGFP	DT4_fuGFP	N/A	BBF10K_003454
DT4_GS flexible linker	DT4_GS flexible linker	N/A	BBF10K_003455
DT5_TEVcut	DT5_TEVcut	N/A	BBF10K_003457
DF_TEVcut_His_Stop_TZ	DF_TEVcut_His_Stop_TZ	N/A	BBF10K_003459
DF_TEVcut_fuGFP_His_Stop_TZ	DF_TEVcut_fuGFP_His_Stop_TZ	N/A	BBF10K_003460
T4T5_Int_ThiolS	T4T5_Int_ThiolS	N/A	BBF10K_003461
T4T5_Int_SaltS	T4T5_Int_SaltS	N/A	BBF10K_003462
T4T5_AmilCP	T4T5_AmilCP	N/A	BBF10K_003463
T4T5_fuGFP	T4T5_fuGFP	N/A	BBF10K_003464
T4T5_GS flexible	T4T5_GS flexible	N/A	BBF10K_003465
T4E_3xStop	T4E_3xStop	N/A	BBF10K_003466
T5E_6H	T5E_6H	N/A	BBF10K_003467
T5E_Calm	T5E_Calm	N/A	BBF10K_003468
T5E_CBD	T5E_CBD	N/A	BBF10K_003469
T5E_Cex	T5E_Cex	N/A	BBF10K_003470
T5E_MBP	T5E_MBP	N/A	BBF10K_003471
T5E_R5	T5E_R5	N/A	BBF10K_003472
T5E_CipA	T5E_CipA	N/A	BBF10K_003473
T5E_3xStop	T5E_3xStop	N/A	BBF10K_003474
EF_ECK9600	EF_ECK9600	N/A	BBF10K_003475
EF_T7term	EF_T7term	N/A	BBF10K_003476
EF_TZ	EF_TZ	N/A	BBF10K_003477

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
Kefentse Arnold Tumedi	tumediak {at} yahoo {dot} com	Botswana
Ery Fukushima	ery.fukushima {at} ikiam {dot} edu {dot} ec	Ecuador
Dong Nguyen Tam	NA	Vietnam
Vitaliy Strochkov	NA	Kazakhstan
Erman ÖZER	aracnoid {at} gmail {dot} com	Turkey
Ferdinand Molnár	NA	Kazakhstan
Emre Yörük	unalsergen {at} gmail {dot} com	Turkey
Shree Ram Singh	singhlabasu {at} gmail {dot} com	United States
Esteban Erben	eerben {at} iib {dot} unsam {dot} edu {dot} ar	Argentina
Juan Vicente Farizano	juan.farizano {at} fbqf {dot} unt {dot} edu {dot} ar	Argentina
Ahmed Hegazy	a.hegazy {at} biotec {dot} rwth-aachen {dot} de	Germany
Ben Thuronyi	bwt2 {at} williams {dot} edu	United States
TING-WEI LIN	weitinglin66 {at} gmail {dot} com	Taiwan
Jennifer Hsieh	j.hsieh {at} kurabiotech {dot} com	Chile
Matthew Phanchana	matthew.pha {at} mahidol {dot} edu	Thailand
Simran Kushwaha	https://www.bits-pilani.ac.in/pilani/biologicalScience/Faculty	India
Ahmed Atef	ahmed_atefaig2 {at} yahoo {dot} com	Saudi Arabia
Emin Bursa	NA	United States
Swaraj Kunal	swaraj.avisa {at} gmail {dot} com	India
Keoni Gandall	koeng101 {at} gmail {dot} com	United States
semih cagan	semihcgn {at} gmail {dot} com	Turkey
James Altamirano	James_altam {at} yahoo {dot} com	United States
Rodrigo Caroca	rcaroca {at} uazuay {dot} edu {dot} ec	Ecuador
Jordan Gonzalez	jgonzalez {at} thecitizensciencelab {dot} org	United States
John Allan	john.allan {at} northumbria {dot} ac {dot} uk	United Kingdom
Jimmy Linhares	Synbioamazonas {at} gmail {dot} com	Brazil
Amjed Alsultan	amjed.talib {at} qu {dot} edu {dot} iq	Iraq
Majed Alghamdi	mafa2 {at} le {dot} ac {dot} uk	United Kingdom
Han Teng Wong	wong_han_teng {at} imcb {dot} a-star {dot} edu {dot} sg	Singapore
Tim Dobbs	tim.dobbs {at} cri-paris {dot} org	France
Aleksandr Shilovich	mercurialbadger {at} yandex {dot} ru	Russia
Kenneth Frimpong	ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh	Ghana
Pratik Vyas	iampratikvyas {at} gamil {dot} com	India
Amitabha Majumdar	Mamitava {at} nccs {dot} res {dot} in	India
Peer Schenk	Reuben.brown {at} uq {dot} edu {dot} au	Australia

Download all of this information as a CSV from our GitHub.

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Use: The Recipient shall use, store, and dispose of the Material and any Modifications in accordance with good laboratory practice and shall ensure compliance with all applicable laws, rules, and regulations, including laws, rules, and regulations governing export control and safety.
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DEFINITIONS:

Material: Original Material, Progeny, and Unmodified Derivatives. The Material shall not include: (a) Modifications, or (b) other substances created by the Recipient through the use of the Material, which are not Modifications, Progeny, or Unmodified Derivatives.
Progeny: Unmodified descendant from the Material, such as virus from virus, cell from cell, or organism from organism.
Unmodified Derivatives: Substances created by the Recipient which constitute an unmodified functional subunit or product expressed by the Original Material. Some examples include: subclones of unmodified cell lines, purified or fractionated subsets of the Original Material, proteins expressed by DNA/RNA supplied by Provider, or monoclonal antibodies secreted by a hybridoma cell line.
Modifications: Substances created by the Recipient which contain/incorporate the Material.