E. coli Protein Expression Toolkit

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
  • 2 destination vectors with pink or blue color screening.

All genes are present on the plate in E. coli glycerol stocks.

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.


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_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_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_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


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
TING-WEI LIN weitinglin66 {at} gmail {dot} com
Ahmed Hegazy a.hegazy {at} biotec {dot} rwth-aachen {dot} de
Rodrigo Caroca rcaroca {at} uazuay {dot} edu {dot} ec
Jordan Gonzalez jgonzalez {at} thecitizensciencelab {dot} org
Vitaliy Strochkov NA
Amitabha Majumdar Mamitava {at} nccs {dot} res {dot} in
Kenneth Frimpong ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh
semih cagan semihcgn {at} gmail {dot} com
Matthew Phanchana matthew.pha {at} mahidol {dot} edu
Amjed Alsultan amjed.talib {at} qu {dot} edu {dot} iq
Keoni Gandall koeng101 {at} gmail {dot} com
James Altamirano James_altam {at} yahoo {dot} com
Ahmed Atef ahmed_atefaig2 {at} yahoo {dot} com
Dong Nguyen Tam NA