Team:Kent/Composite Part

Composite Parts


Bba_K1985007 pSB1A3-[AraC-pBAD]-mamO

Figure 1. Shows part BBa_1985007 in pSB1A3. The image was created using Snapgene. This part is a combination of biobricks BBa_K1985016 and BBa_K1985006. It was used to build further composite parts.



Bba_K1985017 pSB1A3-[AraC-pBAD]-MamOP

Figure 2. Shows part BBa_1985017 in pSB1A3, with the parts assembled in order of ligation. The image was created using Snapgene. This part is a combination of biobricks BBa_K1985016, BBa_K1985006 and BBa_K1985000. It was used to build further composite parts.



Bba_K1985008 pSB1A3-[AraC-pBAD]-MamOPX

Figure 3. Shows part BBa_1985008 in pSB1A3, with the parts assembled in order of ligation. The image was created using Snapgene. This part is a combination of biobricks BBa_K1985016 , BBa_K1985006, BBa_K1985000 and BBa_K1985001. It was used to build further composite parts.



Bba_K1985009 pSB1A3-[AraC-pBAD]-MamOPXT

Figure 4. Shows part BBa_1985009 in pSB1A3, with the parts assembled in order of ligation. The image was created using Snapgene. This part is a combination of biobricks BBa_K1985016 , BBa_K1985006, BBa_K1985000 and BBa_K1985001. This is the BioBrick device construct, which contains four protein coding genes and an arabinose inducible promoter. The MamO protein should nucleate the magnetite crystals [2] allowing the electron transport complex of mamPXT to further develop the magnetite. The part was used in pSB1A3 rather than pSB1C3 as it was cotransformed with pec86, a cytochrome maturation factor, which was in a chloramphenicol resistant plasmid. Pec86 is a pACYC184 derivative containing the E.coli genes, ccmABCDEFGH, of the aeg operon, expressed from the tet promoter of the plasmid. The genes are essential for maturation of cytochromes c, i.e. for the covalent attachment of the heme to the protein [1].



Bba_K1985013 pSB1A3-[AraC-pBAD]-[CsgA-SS-Sup35-1-61]

Figure 5. Shows part BBa_1985013 in pSB1A3. The image was created using Snapgene. This part is an improved version of a previously designed BioBrick (Part:BBa_K1739002), which was designed by the Kent 2015 iGEM team. This part contains three segments: the promoter, the CsgA signal sequence and the first 61 residues of the prion domain, Sup35. Our improved BioBrick aims to optimize the self assembly process of amyloid fibrils with the addition of these 61 residues, as they are considered to be a suitable building block for the assembly of nanostructures. This was inserted into the pSB1A3 backbone and uses the promoter BBa_K1985016, which allows for tighter control of the expression of amyloid fibrils over the previous constitutive promoter.



Bba_K1985014 pSB1A3-[AraC-pBAD]-[CsgA-SS-Sup35-1-61-Cytb562]

Figure 6. Shows part BBa_1985014 in pSB1A3. The image was created using Snapgene. This part is an improved version of the Envirowire fusion protein(BBa_K1739003). It contains 4 segments: the promoter, the CsgA signal sequence, the first 61 residues of the prion forming domain, Sup35 and the electron transfer protein cytochrome 562. Our improved BioBrick aims to optimize the self assembly process of amyloid fibrils with the addition of these 61 residues, as they are considered to be an improved building block for the assembly of nanostructures. It was used in a pSB1A3 backbone and contains part BBa_K1985016, an arabinose inducible promoter.



Bba_K1985015 pSB1A3-[AraC-pBAD]-[CsgA-SS-Sup35NM]

Figure 7. Shows part BBa_1985015 in pSB1A3. The image was created using Snapgene. This part is an improved version of a previously designed BioBrick (Part:BBa_K1739002), which was designed by the Kent 2015 iGEM team. The promoter BBa_J23104 has been removed and instead the part BBa_K1895016 is used, which allows for greater control over the expression of sup35NM. This part contains two segments in addition to the promter; the CsgA signal sequence and Sup35NM. It was used in pBS1A3.





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