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.

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
CE_meffBlue/Rtms5/ NF pocilloporin /Biobrick K1033902	CE_meffBlue/Rtms5/ NF pocilloporin /Biobrick K1033902
CE_sarcGFP	CE_sarcGFP
CE_beta-galactosidase	CE_beta-galactosidase
AF_lacZ pDest	AF_lacZ pDest	N/A
AE_lacZ pDest	AE_lacZ pDest	N/A
AF_high copy (pUC) ori/KanR	AF_high copy (pUC) ori/KanR	N/A
AF_medium copy (pET) ori/KanR	AF_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
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_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_RNAseInhibitor	CD_RNAseInhibitor
CD_gp32	CD_gp32
CD_BsuLF	CD_BsuLF
CD_UvsY	CD_UvsY
CD_UvsX	CD_UvsX
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_TEVcut	DN5_TEVcut	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
AF_spisPink pDest	AF_spisPink pDest	N/A

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