Team:Washington/Magnetosomes/Parts

From 2011.igem.org

The iGEM toolkits group submitted a total twenty-two parts to the registry:

• 10 magnetosome gene groups from mamAB operon
• 3 superassemblies of the mamAB operon from the set of 10 gene groups
• 2 sfGFP-mamK and sfGFP-mamI gene fusions
• 5 Gibson Assembly (pGA) vectors

Magnetosome Toolkit Favorite Parts

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590015 BBa_K590015 sfGFP_mamK_pGA1C3]

This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamK was previously reported to be essential for proper magnetosome chain alignment in magnetotactic bacteria; its bacterial actin-like cytoskeleton protein is required for proper alignment of the magnetosomes in a chain. Beyond magnetosome alignment, its filament could act as a scaffold for protein localization or to alter cell morphology.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590016 BBa_K590016 sfGFP_mamI_pGA1C3]

This part consists of the mamI gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamI is a membrane-localized protein that is essential for magnetosome vesicle formation, and is known to also bind the MamK filament.

The Gibson Assembly Toolkit

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590010 BBa_K590010 pGA1A3_pLacGFP]

This is a high copy plasmid backbone (replication origin pMB1) that confers ampicillin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB1A3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590011 BBa_K590011 pGA1C3_pLacGFP]

This is a high copy plasmid backbone (replication origin pMB1) that confers chloramphenicol resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB1C3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590014 BBa_K590014 pGA3K3_pLacGFP]

This is a medium copy plasmid backbone (replication origin p15A) that confers kanamycin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. pGA3K3 is identical to pSB3K3 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590012 BBa_K590012 pGA4C5_pLacGFP]

This is a low copy plasmid backbone (replication origin pSC101) that confers chloramphenicol resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB4C5 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K590013 BBa_K590013 pGA4A5_pLacGFP]

This is a low copy plasmid backbone (replication origin pSC101) that confers ampicillin resistance. It has an insert with a LacI-repressible promoter driving GFP expression. It is identical to pSB4A5 except for its bglBrick prefix and suffix, and is optimized for Gibson Assembly. It is also notable that though we searched in multiple BioBrick distributions from different years, we could not find any that could be sequence-verified as pSB4A5 (the Vf2 sequence reads all suggested pSB1A2). We then generated pGA4A5 from pGA4C5 using the ampicillin cassette from pSB1A3.