LOCUS       sequence-431872-        6428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tNCS to the peroxisome for toxicity assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6428)
  AUTHORS   Baker JJ, Shi J, Wang S, Mujica EM, Bianco S, Capponi S, Dueber JE
  TITLE     ML-enhanced peroxisome capacity enables compartmentalization of 
            multienzyme pathway.
  JOURNAL   Nat Chem Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2.
  PUBMED    39402374
REFERENCE   2  (bases 1 to 6428)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2024 Oct 14. doi: 10.1038/s41589-024-01759-2."
COMMENT     Sequence Label: pJJB1426
FEATURES             Location/Qualifiers
     source          1..6428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..680
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     550..573
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1185..1895
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     promoter        2411..2598
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2599..3261
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      3476..4458
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4516..5331)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4516..5331)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5239..5258
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5323..5342
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5556..6144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6045..6064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagttcgagt ttatcattat caatactgcc atttcaaaga atacgtaaat aattaatagt
       61 agtgattttc ctaactttat ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt
      121 acatgcccaa aatagggggc gggttacaca gaatatataa catcgtaggt gtctgggtga
      181 acagtttatt cctggcatcc actaaatata atggagcccg ctttttaagc tggcatccag
      241 aaaaaaaaag aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt
      301 ccattctctt agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac
      361 ctcaatggag tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc
      421 atgtatctat ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa
      481 agctgaaaaa aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt
      541 atataaagac ggtaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat
      601 tctactttta tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata
      661 aacacacata aacaaacaaa agatctatgg aagaaactgt aatgttatat cacgaattgg
      721 aagtcgcagc ctcagctgac gaagtatggt ctgtcgaagg ttctcctgaa ttaggtttac
      781 atttaccaga tttgttacca gctggaatat tcgcaaagtt tgagataact ggagatggtg
      841 gagaaggttc tattttggac atgacatttc cacctggtca attccctcat cattatagag
      901 agaagttcgt tttcttcgac cataagaata gatataagtt ggttgaacaa attgacggtg
      961 atttctttga tttgggagta acctactaca tggacactat tagggtcgtc gcaacaggtc
     1021 ctgattcttg tgtcatcaaa tcaacaactg aatatcatgt taaaccagaa tttgctaaga
     1081 ttgttaaacc tttaatcgat acagttccat tagccattat gtcagaggca atagctaagg
     1141 tagtattgga aaataagcac aaatcttctg agggatccgg gtcctctaaa ggtgaagaat
     1201 tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat
     1261 tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattga
     1321 tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg
     1381 gtttgcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg
     1441 ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca
     1501 agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag
     1561 gtattgattt taaagaaggt ggtaacattt taggtcacaa attggaatac aactataact
     1621 ctcacaatgt ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa
     1681 ttagacacaa cattgaagat ggtggtgttc aattagctga ccattatcaa caaaatactc
     1741 caattggtga tggtccagtc ttgttaccag acaaccatta cttatcctat caatctgcct
     1801 tatccaaaga tccaaacgaa aagagagatc acatggtctt gttagaattt gttactgctg
     1861 ctggtattac ccatggtatg gatgaattgt acaaagggtc cttgggaaga ggtagaagat
     1921 ccaaattgta ataatggcag tgcttttaac taagaattat tagtcttttc tgcttatttt
     1981 ttcatcatag tttagaacac tttatattaa cgaatagttt atgaatctat ttaggtttaa
     2041 aaattgatac agttttataa gttacttttt caaagactcg tgctgtctat tgcataatgc
     2101 actggaaggg gaaaaaaaag gtgcacacgc gtggcttttt cttgaatttg cagtttgaaa
     2161 aatgctggaa atctgctcgt cagtggtgct cacactgacg aatcatgtac agatcatacc
     2221 gatgactgcc tggcgactca caactaagca agacagccgg aaccagcgcc ggcgaacacc
     2281 actgcatata tggcatatca caacagtcca cgtctcaagc agttacagag atgttacgaa
     2341 ccactagtgc actgcagtac aaacacagtc ctttcccgca attttctttt tctattactc
     2401 ttggcctcct ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt
     2461 ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa
     2521 ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca
     2581 agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa
     2641 accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc
     2701 ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac
     2761 gtccacacag gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc
     2821 tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgaa
     2881 gactgcggga ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta
     2941 aaaaggtttg gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggttgat
     3001 ctttcgaaca ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggtgat
     3061 ctctcttgcg agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc
     3121 ctccacgttg attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag
     3181 gctcttgcgg ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc
     3241 accaaaggtg ttcttatgta gtgacaccga ttatttaaag ctgctgcata cgatatatat
     3301 acatgtgtat atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac
     3361 tgaagatgac aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgctttcc
     3421 ttttttcttt ttgctttttc tttttttttc tcttgaactc gacggatcat agagtaacga
     3481 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     3541 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     3601 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     3661 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     3721 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     3781 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     3841 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     3901 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     3961 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     4021 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     4081 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     4141 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     4201 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     4261 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     4321 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     4381 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     4441 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaacca
     4501 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     4561 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4621 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4681 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4741 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     4801 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     4861 attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt
     4921 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4981 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5041 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5101 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5161 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5221 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5281 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
     5341 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     5401 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     5461 tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga
     5521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5881 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5941 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6001 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6061 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6121 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6181 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6241 cgtgcggccg cccctgaatt cgcatctaga ctgatgagac gtggtagagc cacaaacagc
     6301 cggtacaagc aacgatctcc aggaccatct gaatcatgcg cggatgacac gaactcacga
     6361 cggcgatcac agacattaac ccacagtaca gacactgcga caacgtggca attcgtcgca
     6421 atacaacg
//
