PARTS

BBa_k2223000: 4abh

The 4abh gene, native to A. bisporus, allows the conversion of molecules like anthranilate and para-aminobenzoic acid to 4-aminophenol.

BBa_k2223001: nhoA

The nhoA gene is native to E. coli and allows for the conversion of 4-aminophenol to acetaminophen.

The enzyme created by the gene ssuE catalyzes activity that reduces the Flavin mononucleotide (FMN) by using NADH dehydrogenase as a reducing agent. This gene sequenced was optimized from S. elonagtus PCC 7002 sequence.

BBa_k2223003: bluB

The gene bluB encodes an enzyme that catalyzes an oxidized reaction that cleaves the reduced Flavin mononucleotide (FMNH₂) transforming to 5,6-dimethylbenzimidazole (5,6-DMB). The bluB sequence was optimized for PCC 7942 using the reference genome sequence from Sinorhizobium meliloti 1021.

BBa_k2223004: riboswitch

The riboswitch part contains a psbAI promoter which transcribes an mRNA strand consisting of a riboswitch and repressor gene, TetR. The riboswitch binds to vitamin B₁₂ and regulates the expression of the downstream repressor gene. In the presence of vitamin B₁₂, the riboswitch specifically anneals onto the cobalamin molecule inducing a conformational change within the mRNA sequestering the riboswitch binding site upstream of TetR. However, in the absence of vitamin B₁₂, TetR is continuously expressed.

A light-induced psbAI (Part: BBa_K1913011) promoter, documented to be constitutively expressed in cyanobacteria such as S. elongatus PCC 7942, was modified at the -10 element for expression in E. coli. The promoter was placed upstream of the biobrick parts, producing two separate mRNA strands. The reporter for this part is in BBa_K2223005.

BBa_k2223005: riboswitch_reporter

The riboswitch_reporter part contains a psbAI promoter which transcribes an mRNA strand consisting of a Tet operator, TetO, upstream of the reporter gene, mRFP. The reporter is trans-regulated by a Tet repressor protein bound to the Tet operator hindering translation. In the presence of vitamin B₁₂ the Tet systems renders useless thus, mRFP is expressed.

A light-induced psbAI (Part:BBa_K1913011) promoter, documented to be constitutively expressed in cyanobacteria such as S. elongatus PCC 7942, was modified at the -10 element for expression in E. coli. The promoter was placed upstream of the biobrick parts, producing two separate mRNA strands. The riboswitch paired with this reporter is BBa_K2223004.