Overview

Formosa’s parts are labeled BBa_K5437000 to BBa5437020. Our plasmids were designed to express many enzymes, meaning that basic parts will code for individual proteins. In contrast, composite parts will code for the overexpressed or controlled expressed protein groups.

Basic Parts

Our basic parts are mainly comprised of the individual genes of the MVA, MEP, GPP synthesis, and L-borneol synthesis pathways. We created new parts because of our novel optimization strategies which altered the sequences to fit our strains of E. coli expression.

Part Number Part Name Description Type Length
BBa_K3633015 T7 Promoter Strong constitutive promoter for overexpression of key enzymes Promoter 19 bp
BBa_K4491007 araBAD promoters Inducible promoter for lower and controlled expression Promoter 166 bp
BBa_K4150006 T7 terminator Efficient transcription termination in pBAD plasmid Terminator 129 bp
BBa_K5437001 pACYC-LBB DXS DXP Synthase (DXS) which turns G3P and pyruvate into 1-Deoxyxylulose-5-phosphate synthase (DXP) DNA 1860 bp
BBa_K5437002 pACYC-LBB DXR DXP reductoisomerase (DXR) converts 1-Deoxy-D-xylulose-5-phosphate (DXP) into methylerythritol phosphate (MEP) DNA 1194 bp
BBa_K5437003 pACYC-LBB IDI Isopentenyl diphosphate isomerase (IDI) stabilizes the levels of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) DNA 1050 bp
BBa_K5437004 pACYC-LBB IspA IspA codes for farnesyl diphosphate synthase that turns IPP and DMAPP into geranyl diphosphate (GPP) and farnesyl diphosphate (FPP) DNA 897 bp
BBa_K5437005 pACYC-LBB BbTPS3 L-bornyl diphosphate synthase (BbTPS3) turns geranyl diphosphate into bornyl diphosphate DNA 1668 bp
BBa_K5437006 pACYC-LBB ALP Alkaline phosphatase dephosphorylates L-bornyl diphosphate into L-borneol DNA 1416 bp
BBa_K5437007 pACYC-LBB HmgR 3-hydroxy-3-methyl-glutaryl-coenzyme A (Hmg-CoA) reductase converts Hmg-CoA into mevalonate DNA 3135 bp
BBa_K5437009 pBAD-LBB ACCT Coding sequence for acetoacetyl-CoA thiolase (ACCT enzyme) converting Acetyl-CoAe to Acetoaceyl-CoA. DNA 1191 bp
BBa_K5437025 pBAD-LBB HmgS Coding sequence for 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase (HMGS enzyme) converting HMG-CoA to Mevalonate. DNA 1476 bp
BBa_K5437011 pBAD-LBB MK Mevalonate kinase converts mevalonate into mevalonate-5-phosphate (MVAP) DNA 1332 bp
BBa_K5437012 pBAD-LBB PMK MVAP kinase converts mevalonate-5-phosphate to mevalonate-5-diphosphate DNA 1356 bp
BBa_K5437013 pBAD-LBB MDD MVAPP decarboxylase converts mevalonate-5-diphosphate (MVAPP) into isopentenyl diphosphate DNA 1191 bp
BBa_K5437014 pBAD-LBB IspD Coding sequence for 4-diphosphocytidyl-2C-methyl-d-erythritol (CDP-ME) cytidylyltransferase (IspD enzyme) converting MEP to CDP-ME. DNA 711 bp
BBa_K5437015 pBAD-LBB IspE Coding sequence for CDP-ME kinase (IspE enzyme) converting CDP-ME to CDP-MEP. DNA 852 bp
BBa_K5437016 pBAD-LBB IspF Coding sequence for 2C-methyl-d-erythritol-2,4-cyclo-diphosphate (MEC) synthase (IspF enzyme) converting CDP-MEP to MEC. DNA 480 bp
BBa_K5437017 pBAD-LBB IspG Coding sequence for 4-hydroxy-3-methyl-2-(E)-butenyl-4-diphosphate (HMBPP) synthase (IspG enzyme) converting MEC to HMBPP. DNA 1119 bp
BBa_K5437018 pBAD-LBB IspH IspH converts 4-hydroxy-3-methylbut-2-enyl diphosphate (HMBPP) into isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) DNA 951 bp

Composite Parts

Our composite parts are the sections responsible for gene expression within our plasmids pACYC-LBB and pBAD-LBB, with the major difference being the order of expression that each composite part is capable of.

Part Number Part Name Description Type Length
BBa_K5437019 Overexpressed MCS Overexpression proteins for original rate-limiting steps Composite 11860 bp
BBa_K5437022 Controlled Express MCS Controlled expression proteins for non-rate-limiting steps Composite 11850 bp