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.

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
BT1_BCD2	BT1_BCD2	N/A
BT1_Riboj54_F1	BT1_Riboj54_F1	N/A
BT1_Riboj_B34	BT1_Riboj_B34	N/A
BT1_BCD12	BT1_BCD12	N/A
BT1_BCD23	BT1_BCD23	N/A
BT1_BCD2_His	BT1_BCD2_His	N/A
BT1_RiboJ_B34_His	BT1_RiboJ_B34_His	N/A
BT1_BCD2_DsbA	BT1_BCD2_DsbA	N/A
BT1_BCD2_OmpT	BT1_BCD2_OmpT	N/A
BT1_BCD2_pelB	BT1_BCD2_pelB	N/A
T1T2_Calm	T1T2_Calm	N/A
T1T2_CBD	T1T2_CBD	N/A
T1T2_Cex	T1T2_Cex	N/A
T1T2_MBP	T1T2_MBP	N/A
T1T2_R5	T1T2_R5	N/A
T1T2_GST	T1T2_GST	N/A
T1T2_CipA	T1T2_CipA	N/A
T1T2_S-Tag	T1T2_S-Tag	N/A
T1T2_GB1	T1T2_GB1	N/A
T1T2_GS flexible	T1T2_GS flexible	N/A
T1T3_GS flexible	T1T3_GS flexible	N/A
T1C_GS flexible	T1C_GS flexible	N/A
T1C_His_TEVcut	T1C_His_TEVcut	N/A
BT2_BCD2_His	BT2_BCD2_His	N/A
BT2_RiboJ_B34	BT2_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
T2C_GS flexible	T2C_GS flexible	N/A
T2C_TEVcut	T2C_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
T2T3_int_∆I	T2T3_int_∆I
T2T3_SUMO	T2T3_SUMO
T2T3_fuGFP	T2T3_fuGFP
T2T3_AmilCP	T2T3_AmilCP
T2T3_GS flexible	T2T3_GS flexible
T3C_SUMO	T3C_SUMO
T3C_TEV	T3C_TEV
T3C_int_∆I	T3C_int_∆I
T3C_fuGFP	T3C_fuGFP
T3C_GS flexible linker	T3C_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
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
DT4_Int_SaltS	DT4_Int_SaltS	N/A
DT4_Int_ThiolS	DT4_Int_ThiolS	N/A
DT4_TEV	DT4_TEV	N/A
DT4_fuGFP	DT4_fuGFP	N/A
DT4_GS flexible linker	DT4_GS flexible linker	N/A
DT5_TEVcut	DT5_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
T4T5_Int_ThiolS	T4T5_Int_ThiolS	N/A
T4T5_Int_SaltS	T4T5_Int_SaltS	N/A
T4T5_AmilCP	T4T5_AmilCP	N/A
T4T5_fuGFP	T4T5_fuGFP	N/A
T4T5_GS flexible	T4T5_GS flexible	N/A
T4E_3xStop	T4E_3xStop	N/A
T5E_6H	T5E_6H	N/A
T5E_Calm	T5E_Calm	N/A
T5E_CBD	T5E_CBD	N/A
T5E_Cex	T5E_Cex	N/A
T5E_MBP	T5E_MBP	N/A
T5E_R5	T5E_R5	N/A
T5E_CipA	T5E_CipA	N/A
T5E_3xStop	T5E_3xStop	N/A
EF_ECK9600	EF_ECK9600	N/A
EF_T7term	EF_T7term	N/A
EF_TZ	EF_TZ	N/A

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
Rodrigo Caroca	rcaroca {at} uazuay {dot} edu {dot} ec
Vitaliy Strochkov	NA
Amitabha Majumdar	Mamitava {at} nccs {dot} res {dot} in
Dong Nguyen Tam	NA
Amjed Alsultan	amjed.talib {at} qu {dot} edu {dot} iq
Matthew Phanchana	matthew.pha {at} mahidol {dot} edu
Jordan Gonzalez	jgonzalez {at} thecitizensciencelab {dot} org
Keoni Gandall	koeng101 {at} gmail {dot} com
semih cagan	semihcgn {at} gmail {dot} com

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MTA

Unilateral OpenMTA

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