LOCUS       sequence-432265-        7596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Pex32.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  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 7596)
  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: pJJB1336
FEATURES             Location/Qualifiers
     source          1..7596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1964..2151
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2019..2043)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2627..3034
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3035..4129
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(5160..5975)
                     /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(5160..5975)
                     /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     5883..5902
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5967..5986
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6200..6788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6689..6708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagaggatgt ccaatatttt ttttaaggaa taaggatact tcaagactag attcccccct
       61 gcattcccat cagaaccgta aaccttggcg ctttccttgg gaagtattca agaagtgcct
      121 tgtccggttt ctgtggctca caaaccagcg cgcccgatat ggctttcttt tcacttatga
      181 atgtaccagt acgggacaat tagaacgctc ctgtaacaat ctctttgcaa atgtggggtt
      241 acattctaac catgtcacac tgctgacgaa attcaaagta aaaaaaaatg ggaccacgtc
      301 ttgagaacga tagattttct ttattttaca ttgaacagtc gttgtctcag cgcgctttat
      361 gttttcattc atacttcata ttataaaata acaaaagaag aatttcatat tcacgcccaa
      421 gaaatcaggc tgctttccaa atgcaattga cacttcatta gccatcacac aaaactcttt
      481 cttgctggag cttcttttaa aaaagacctc agtacaccaa acacgttacc cgacctcgtt
      541 attttacgac aactatgata aaattctgaa gaaaaaataa aaaaattttc atacttcttg
      601 cttttattta aaccattgaa tgatttcttt tgaacaaaac tacctgtttc accaaaggaa
      661 atagaaagaa aaaatcaatt agaagaaaac aaaaaacaaa agatctatgg acacaaattc
      721 taaaaccaaa gtacaaactg aaaataaaaa aattaaggcg aaatttatac ataatcatgg
      781 gcaaaagccc tctcttatcc aaattacacc tccaatgatt tccagcactc tgtttcacgc
      841 atatccacta ctactaatat tcgacaatgc cttagcgaat ataatgtggc tgtccgacga
      901 taaatgttta acattcattt atctaacgag catatggctg accataagtt tttttatccc
      961 cgttgaaaca gaggctagcc attttctgcc atttacaaag attttgagac tatggttagg
     1021 cattataagc ggggcatttc tattcttatc attcatgtat tatattgttt cattgatagc
     1081 ttcgttaaga gacacggaac ctcctacatt ggatgaaatt gtcgtgctac tagagtctgt
     1141 gttggataaa ctggaagtac tgagaaacga gctgaatgtt tggaaaaaat taaagctttc
     1201 atttgatggc gtcaataagg agtgttctgg caagaggttg ttttgcagat tattcttatt
     1261 cggtacaatt tttcaaatta ttatcatgag atacatatca ccgggaactt atactagatt
     1321 ttttattatt accggcttga tatacaatac gagtagtttt caagctacct taaggctact
     1381 ttggaggttt actgctgtta ggaatttcta ttatttgggg attgaaagtt ttaaaatttc
     1441 gagtttctta ccaaaacact taaagatgga acaaattatt cctttatctc aaggacgtgc
     1501 aatcacggtg ccactggtag aggtattgcc taaactactt cgggacaaga agggtgatga
     1561 ccacattcat attctacagc ttctacttaa tgagcaaaaa gataactttg gtaatgaaga
     1621 ccttaaaata ctagaaattg aagtatacga aaatcaaaga aggtggtatc aaaataaaaa
     1681 ttggagtaca aaattactgc cgtacgaaag acaaaactac tgcatagaaa ttaaaaacac
     1741 tgatggaact ttaaccatga ggagttgttt gcccccagat ggacttggtg aagaagaatt
     1801 acccaataat tggcattgga tcaatgataa ttgggacgga actgattgga tatattcaga
     1861 ttcagcatgg aaagaaattg gacaatacag ttctttggaa agttttacca ggtcaagaaa
     1921 atggaaacga cgcctcttcc atttgtaagg atcctaactc gaggcgaatt tcttatgatt
     1981 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2041 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2101 gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccgag
     2161 caaatgcctg caaatcgctc cccatttcgc tggaaatctg ctcgtcagtg gtgctcacac
     2221 tgacgaatca tgtacagatc ataccgatga ctgcctggcg actcacaact aagcaagaca
     2281 gccggaacca gcgccggcga acaccactgc atatatggca tatcacaaca gtccacgtct
     2341 caagcagtta cagagatgtt acgaaccact agtgcactgc agtacatcga ggagaacttc
     2401 tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat
     2461 caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa
     2521 aaacgttata tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc
     2581 cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac
     2641 tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac
     2701 ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt
     2761 tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa
     2821 aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta
     2881 aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc
     2941 aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat
     3001 atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt
     3061 tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc
     3121 tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc
     3181 tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct ccaagaaggc
     3241 tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggaaccggta gtgttagacc
     3301 tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg
     3361 taactttgca tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg
     3421 tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga
     3481 agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag
     3541 aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc
     3601 cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac
     3661 catcaagaac gaatttccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat
     3721 gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg
     3781 tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc
     3841 gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg
     3901 ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc
     3961 aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt
     4021 taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac
     4081 cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct
     4141 ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa
     4201 attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt
     4261 tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg
     4321 gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc
     4381 accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat
     4441 tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac
     4501 catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc
     4561 ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgagagt actcgtatcg
     4621 catgtcggtg cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaaactata
     4681 tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag gaatacaggt
     4741 aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa
     4801 tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta
     4861 tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa
     4921 aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc
     4981 ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct taacgacgta
     5041 gtcgacctcc tcttcagtac taaatctacc aataccaaat ctgatggaag aatgggctaa
     5101 tgcatcatcc ttacccagcg ccgagcggcc gcgtgttaca accaattaac caattctgat
     5161 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     5221 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     5281 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     5341 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     5401 gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag
     5461 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     5521 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     5581 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     5641 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     5701 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     5761 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     5821 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     5881 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     5941 ctggagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     6001 taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag
     6061 agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat
     6121 cagtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6841 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtgcg gccgccaata
     6901 cccagaagaa cagtaaaata aagcaaggta cgtgaaatta atatttttaa atggttctaa
     6961 ccgatgccga agaactgcgc agtccggtta taacgtctga catgtccttt tttgatttgg
     7021 aatccaacca ctcaagtgac tctgttcatt tactttgcga aaaatatacc cacaaattgc
     7081 ccatcgaaag tgaatcgcaa accaccttca gactggcacc gacaaagcaa agattataca
     7141 gacagagtac tttatacgta ccgttaagtc tcaagcaaag ggttttctta tttactgaac
     7201 gggtaaagag tatctgggcc ggcttgccaa gatgcaaacc gaataagtat ttcaaagttg
     7261 catttgcctt agccgtcctg acaccattgg ctatttggat attttatatt gactttcgtg
     7321 tacattgatc acatcgactg ttctattggc aaatgaacca cgggcattga ctatttttca
     7381 ggttactact atatattatc atcacgggca aggattgtac cctgaattcg catctagact
     7441 gatgagacgt ggtagagcca caaacagccg gtacaagcaa cgatctccag gaccatctga
     7501 atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc acagtacaga
     7561 cactgcgaca acgtggcaat tcgtcgcaat acaacg
//
