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Composite Parts

Part: BBa_K3268001

Lac-inducible generator of Lpp-OmpA(46-159)-mOR103-15 + His6 tag

This part contains a lac promoter and strong ribosome binding site for lac-inducible expression of the fusion protein of Lpp signal peptide, OmpA aa46-159, and mOR103-15 + His6 tag. This expression should display streptavidin on the cell surface of E. coli.

In order to display mOR103-15 on the surface of cell, we found that the Lpp′OmpA(46-159) hybrid protein can serve as an efficient targeting vehicle for localizing a variety soluble proteins onto the E. coli surface.We got the part BBa_J36850 which is combined with a lac promoter and strong ribosome binding site for lac-inducible expression of the fusion protein of Lpp signal peptide, OmpA aa46-159, and streptavidin wild-type + His6 tag from iGEM kit 2019.

Fig 1. Plasmid map of BBa_K3268001.

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Part: BBa_K3268002

Strong expression of eforRed in E. coli


1. The eforRed coding region was PCR amplified from BBa_K592012 using high-fidelity enzyme KOD-Plus.
2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.
3. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence.
4. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.

Fig 2. Plasmid map of BBa_K3268002.

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Part: BBa_K3268003

Strong expression of AmilCP_Blue in E. coli


1. The AmilCP_Blue coding region was PCR amplified from BBa_K592009 using high-fidelity enzyme KOD-Plus.
2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.
3. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence.
4. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.

Fig 3. Plasmid map of BBa_K3268003.

>> See details on Parts Registry

Part: BBa_K3268004

Strong expression of sfGFP in E. coli


1. The sfGFP coding region was PCR amplified from BBa_K1321337 using high-fidelity enzyme KOD-Plus.
2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.
3. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence.
4. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.

Fig 4. Plasmid map of BBa_K3268004.

>> See details on Parts Registry

Part: BBa_K3268005

Strong expression of mCherry in E. coli


1. The mCherry coding region was PCR amplified from BBa_J18932 using high-fidelity enzyme KOD-Plus.
2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.
3. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence.
4. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.

Fig 5. Plasmid map of BBa_K3268005.

>> See details on Parts Registry