Team:Caltech/Project/Vitamins/Parts

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Folate Production Parts


Basic Parts

{{{{!}}}border="1" {{{!}}}+ Basic Parts (Extracted from L.lactis subspe. IL1403 genome)

Part Name Registry # Description Cloned? Sequence confirmed?

{{{!}}}-

Entire folate synthesis operon

{{{!}}} BBa_K137002{{{!}}}{{{!}}} Includes folB+folKE+folP+ylgG+folC {{{!}}}{{{!}}} NO {{{!}}}{{{!}}} NO {{{!}}}-

folB

{{{!}}} BBa_K137009{{{!}}}{{{!}}} dihydroneopterin aldolase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folKE

{{{!}}} BBa_K137011{{{!}}}{{{!}}} fusion gene: GTP cyclohydrolase & 2-amino-4-hydroxy-6- hydroxymethyldihydropteridine pyrophosphokinase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folP

{{{!}}} BBa_K137012{{{!}}}{{{!}}} Dihydropteroate synthase {{{!}}}{{{!}}} NO{{{!}}}{{{!}}} NO {{{!}}}-

folC

{{{!}}} BBa_K137013{{{!}}}{{{!}}} folate synthetase/polyglutamyl folate synthetase {{{!}}}{{{!}}} NO{{{!}}}{{{!}}} NO {{{!}}}-

pabA

{{{!}}} BBa_K137005{{{!}}}{{{!}}} para-aminobenzoate synthetase component II {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

pabB

{{{!}}} BBa_K137006{{{!}}}{{{!}}} para-aminobenzoate synthetase component I {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}- {{{!}}}}

Construction Intermediates

{{{{!}}} border="1" {{{!}}}+ Construction Intermediates: Adding b0034 (strong RBS) before each individual gene

Part Name Registry # Description Cloned? Sequence confirmed?

{{{!}}}-

folB + b0034

{{{!}}} BBa_S03957{{{!}}}{{{!}}} RBS + dihydroneopterin aldolase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folKE + b0034

{{{!}}} BBa_S03958{{{!}}}{{{!}}} RBS + fusion gene: GTP cyclohydrolase & 2-amino-4-hydroxy-6- hydroxymethyldihydropteridine pyrophosphokinase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folP + b0034

{{{!}}} BBa_S03959{{{!}}}{{{!}}} RBS + Dihydropteroate synthase {{{!}}}{{{!}}} NO{{{!}}}{{{!}}} NO {{{!}}}-

folC + b0034

{{{!}}} BBa_S03960{{{!}}}{{{!}}} RBS + folate synthetase/polyglutamyl folate synthetase {{{!}}}{{{!}}} NO{{{!}}}{{{!}}} NO {{{!}}}-

pabA + b0034

{{{!}}} BBa_S03976{{{!}}}{{{!}}} RBS + para-aminobenzoate synthetase component II {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

pabB + b0034

{{{!}}} BBa_S03977{{{!}}}{{{!}}} RBS + para-aminobenzoate synthetase component I {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

b0034 + folB + b0034 + folKE

{{{!}}} BBa_S03961{{{!}}}{{{!}}} Combining folB (with RBS) + folKE (with RBS) {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}- {{{!}}}}

{{{{!}}} border="1" {{{!}}}+ Construction Intermediates: Adding j23100 (constitutive promoter) to each gene (with RBS already)

Part Name Registry # Description Cloned? Sequence confirmed?

{{{!}}}-

folB + j23100

{{{!}}} BBa_S04032{{{!}}}{{{!}}} Promoter(j23100) + RBS(b0034) + dihydroneopterin aldolase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folKE + j23100

{{{!}}} BBa_S04033{{{!}}}{{{!}}} Promoter + RBS + fusion gene: GTP cyclohydrolase & 2-amino-4-hydroxy-6- hydroxymethyldihydropteridine pyrophosphokinase {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

pabA + j23100

{{{!}}} BBa_S04034{{{!}}}{{{!}}} Promoter + RBS + para-aminobenzoate synthetase component II {{{!}}}{{{!}}} YES{{{!}}}{{{!}}} YES {{{!}}}-

folBKE + j23100

{{{!}}} BBa_S04035{{{!}}}{{{!}}} Promoter + folB (with RBS) + folKE (with RBS) {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} YES {{{!}}}-

pabB + j23100

{{{!}}} BBa_S04039{{{!}}}{{{!}}} Promoter + pabB {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} YES {{{!}}}-

pabB + b0015

{{{!}}} BBa_S04038{{{!}}}{{{!}}} pabB + b0015 terminator {{{!}}}{{{!}}} MAYBE {{{!}}}{{{!}}} WAITING {{{!}}}- {{{!}}}}

Composite Parts

{{{{!}}} border="1" {{{!}}}+ Composite Parts: Adding b0015 (double terminator) to complete constructs with promoter + RBS already

Part Name Registry # Description Cloned? Sequence confirmed?

{{{!}}}-

folB + b0015

{{{!}}} BBa_K137053{{{!}}}{{{!}}} Promoter(j23100) + RBS(b0034) + folB (dihydroneopterin aldolase) + double terminator (b0015) {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} YES {{{!}}}-

folKE + b0015

{{{!}}} BBa_K137054{{{!}}}{{{!}}} Promoter + RBS + folKE (fusion gene: GTP cyclohydrolase & 2-amino-4-hydroxy-6- hydroxymethyldihydropteridine pyrophosphokinase) + double terminator (b0015) {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} YES {{{!}}}-

pabA + b0015

{{{!}}} BBa_K137055{{{!}}}{{{!}}} Promoter + RBS + pabA (para-aminobenzoate synthetase component II) {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} YES {{{!}}}-

pabB + b0015

{{{!}}} BBa_K137109{{{!}}}{{{!}}} Promoter + RBS + pabB (para-aminobenzoate synthetase component I) {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} NO {{{!}}}-

folBKE + b0015

{{{!}}} BBa_K137056{{{!}}}{{{!}}} Promoter + RBS + folB + folKE + b0015 {{{!}}}{{{!}}} YES {{{!}}}{{{!}}} NO {{{!}}}- {{{!}}}}

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