E. coli Protein Expression Tookit

E. coli Protein Expression Tookit

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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:

  • 6 promoters ( T7, constitutive, light inducible, temperature inducible)
  • 5 RBS and insulators
  • 10 purification tags and
  • 6 cleavage sites, flexible linkers
  • 20 genes of which 2 reporters and 18 of genes used in diagnostics
  • two destination vectors with pink or blue color screening.

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.

Where can I find more information:


Check the list of available DNA parts for Expression Toolkit to know which DNA parts are currently available and which are still in the process of being synthesized. Thanks for your patience!


Check out our assembly scheme to see how the DNA parts of this collection fits together.

Genes

Gene Name NCBI ID
AF_lacZ pDest AF_lacZ pDest N/A
AE_spisPink pDest AE_spisPink pDest N/A
AE_lacZ pDest AE_lacZ pDest N/A
high copy (pUC) ori/KanR high copy (pUC) ori/KanR N/A
medium copy (pET) ori/KanR medium copy (pET) ori/KanR N/A
AB_T7 AB_T7 N/A
AB_T7_lacO AB_T7_lacO N/A
AB_pTac AB_pTac N/A
AB_J23100 AB_J23100 N/A
AB_Dawn AB_Dawn N/A
AB_cI875/PL/PR AB_cI875/PL/PR N/A
BN1_BCD2 BN1_BCD2 N/A
BN1_Riboj54_F1 BN1_Riboj54_F1 N/A
BN1_Riboj_B34 BN1_Riboj_B34 N/A
BN1_BCD12 BN1_BCD12 N/A
BN1_BCD23 BN1_BCD23 N/A
BN1_BCD2_His BN1_BCD2_His N/A
BN1_RiboJ_B34_His BN1_RiboJ_B34_His N/A
BN1_BCD2_DsbA BN1_BCD2_DsbA N/A
BN1_BCD2_OmpT BN1_BCD2_OmpT N/A
BN1_BCD2_pelB BN1_BCD2_pelB N/A
N1N2_H6 N1N2_H6 N/A
N1N2_Calm N1N2_Calm N/A
N1N2_CBD N1N2_CBD N/A
N1N2_Cex N1N2_Cex N/A
N1N2_MBP N1N2_MBP N/A
N1N2_R5 N1N2_R5 N/A
N1N2_GST N1N2_GST N/A
N1N2_CipA N1N2_CipA N/A
N1N2_S-Tag N1N2_S-Tag N/A
N1N2_GB1 N1N2_GB1 N/A
N1N2_GS flexible N1N2_GS flexible N/A
N1N3_GS flexible N1N3_GS flexible N/A
N1C_GS flexible N1C_GS flexible N/A
N1C_His_TEVcut N1C_His_TEVcut N/A
BN2_BCD2_His BN2_BCD2_His N/A
BN2_RiboJ_B34 BN2_RiboJ_B34 N/A
BC_B0034 BC_B0034 N/A
BC_B0032 BC_B0032 N/A
BC_RiboJ_RBS64 BC_RiboJ_RBS64 N/A
BC_RiboJ_B34 BC_RiboJ_B34 N/A
BC_BCD23 BC_BCD23 N/A
BC_BCD12 BC_BCD12 N/A
BC_BCD2 BC_BCD2 N/A
N2C_GS flexible N2C_GS flexible N/A
N2C_TEVcut N2C_TEVcut N/A
BC_BCD2_His_TEVcut BC_BCD2_His_TEVcut N/A
BC_BCD2_His_fuGFP_TEVcut BC_BCD2_His_fuGFP_TEVcut N/A
N2N3_int_∆I N2N3_int_∆I
N2N3_SUMO N2N3_SUMO
N2N3_fuGFP N2N3_fuGFP
N2N3_AmilCP N2N3_AmilCP
N2N3_GS flexible N2N3_GS flexible
N3C_SUMO N3C_SUMO
N3C_TEV N3C_TEV
N3C_int_∆I N3C_int_∆I
N3C_fuGFP N3C_fuGFP
N3C_GS flexible linker N3C_GS flexible linker N/A
CD_OMMLV CD_OMMLV
CD_HIVRT CD_HIVRT
CD_Bst-LF CD_Bst-LF
CD_OpenVent CD_OpenVent
CD_PPATho CD_PPATho
CD_RNAseInhibitor CD_RNAseInhibitor
CD_gp32 CD_gp32
CD_BsuLF CD_BsuLF
CD_UvsY CD_UvsY
CD_UvsX CD_UvsX
CE_T7RNApol_Stop CE_T7RNApol_Stop
CD_PBCV-1_ligase CD_PBCV-1_ligase
CD_HRP CD_HRP
CD_TEV_protease CD_TEV_protease
CD_SUMO_protease CD_SUMO_protease
CD_Calmodulin CD_Calmodulin
CD_fuGFP CD_fuGFP
CD_AmilCP CD_AmilCP
CD_TthRNAseH CD_TthRNAseH
CD_EcRNAseH CD_EcRNAseH
DN4_Int_SaltS DN4_Int_SaltS N/A
DN4_Int_ThiolS DN4_Int_ThiolS N/A
DN4_TEV DN4_TEV N/A
DN4_fuGFP DN4_fuGFP N/A
DN4_GS flexible linker DN4_GS flexible linker N/A
DN5_GS flexible DN5_GS flexible N/A
DN5_TEVcut DN5_TEVcut N/A
DE_3xStop DE_3xStop N/A
DF_TEVcut_His_Stop_TZ DF_TEVcut_His_Stop_TZ N/A
DF_TEVcut_fuGFP_His_Stop_TZ DF_TEVcut_fuGFP_His_Stop_TZ N/A
N4N5_Int_ThiolS N4N5_Int_ThiolS N/A
N4N5_Int_SaltS N4N5_Int_SaltS N/A
N4N5_AmilCP N4N5_AmilCP N/A
N4N5_fuGFP N4N5_fuGFP N/A
N4N5_GS flexible N4N5_GS flexible N/A
N4E_3xStop N4E_3xStop N/A
N5E_6H N5E_6H N/A
N5E_Calm N5E_Calm N/A
N5E_CBD N5E_CBD N/A
N5E_Cex N5E_Cex N/A
N5E_MBP N5E_MBP N/A
N5E_R5 N5E_R5 N/A
N5E_CipA N5E_CipA N/A
N5E_3xStop N5E_3xStop N/A
EF_ECK9600 EF_ECK9600 N/A
EF_T7term EF_T7term N/A
EF_TZ EF_TZ N/A
pCO_TZ backbone pCO_TZ backbone N/A
AF_spisPink pDest AF_spisPink pDest N/A
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