LOCUS       sequence-435277-        7539 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express Pex25.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  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 7539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJJB715
FEATURES             Location/Qualifiers
     source          1..7539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1907..2094
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1962..1986)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2570..2977
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2978..4072
                     /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(5103..5918)
                     /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(5103..5918)
                     /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     5826..5845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5910..5929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6143..6731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6632..6651
                     /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 agatctatga gtcagtttgg
      721 cacgacagat atcgtttccg gttcagaaac accaccttat tctggggcaa gttaccagga
      781 cgcccaggat gacaataccc atccacattc atcagacgcc ggtgcagaga agttcagtgc
      841 aggcagtgga agcgagtcgc atactgaatc ctcgagaagt gatgatgagg attctcaagc
      901 aaaaacaaag atggtggata atattaccat cttaaaatat atattggatt ctctatctgg
      961 tagagacaaa ttagccaaaa ttatcaaata cgctctggat atattaaaac tgtttatcga
     1021 aaagtctaag cgaaatttaa ctgttttgga cccttccgta ttaacgtact ataccaagat
     1081 tttgaaaaat ctgactgtaa aagtcgcact aaggcatcca atcacggtaa ttaaagtact
     1141 tctcctgtct cttttgagaa attttgacaa gaaaattgat tttatcagtc aacaattaag
     1201 tacttttaga tatattttaa ggtttggcgg cacaccattc cgagtgtgca gtttcttggg
     1261 gaaattcaat aaaaccagaa agtgcaactt tcaaattgac caaattaaga aaatatggtt
     1321 taatgaagct tcattgagag aatttttgga cttgtattat ggtatctttg atgaattaga
     1381 cttgctatac aaattgaaaa tatggacaaa taaatcattc tattcctttg tcagtagaca
     1441 agagtctctt gcatggcaat acgacattct tttaagcttg aaggaccatt ggctcaacct
     1501 gcaaagctta cagaagagac aattagaatt ggaagtgcaa ctaaaagttc aaaataacgc
     1561 gttactactc tcacccatac tgatgcatca ggcccataaa gacgatggct cacaatcgcc
     1621 aattaggaaa cagttgttaa atgacctaaa tgttaataac gacgcagagg tgttaataca
     1681 caaacaacta aaggccatca aggatgaaaa gacacttgtt tatttagata ttgcaagatt
     1741 atcgtttgat tgcatggcta atacatcaga tattctaaat ttaaaaactc caaagggaac
     1801 gtatgccgtt ctgtccctgg gatccggctt aactggactg gttaaacttt ggataacaac
     1861 aaagaggtca ctttgctctt caaaagattg aggatcctaa ctcgaggcga atttcttatg
     1921 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     1981 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     2041 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc
     2101 gagcaaatgc ctgcaaatcg ctccccattt cgctggaaat ctgctcgtca gtggtgctca
     2161 cactgacgaa tcatgtacag atcataccga tgactgcctg gcgactcaca actaagcaag
     2221 acagccggaa ccagcgccgg cgaacaccac tgcatatatg gcatatcaca acagtccacg
     2281 tctcaagcag ttacagagat gttacgaacc actagtgcac tgcagtacat cgaggagaac
     2341 ttctagtata tctacatacc taatattatt gccttattaa aaatggaatc ccaacaatta
     2401 catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt
     2461 caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa ttggttgttt
     2521 ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa
     2581 tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc
     2641 aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt
     2701 ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga
     2761 aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga
     2821 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
     2881 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
     2941 aatatatata tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt
     3001 cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa
     3061 agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg
     3121 tgctgctatc gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa
     3181 ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggaaccg gtagtgttag
     3241 acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc
     3301 atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa
     3361 aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa
     3421 ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca
     3481 aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg
     3541 gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga
     3601 aaccatcaag aacgaatttc ctacattgaa ggttcaacat caattgattg attctgccgc
     3661 catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt
     3721 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
     3781 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
     3841 cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc
     3901 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
     3961 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
     4021 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc
     4081 tcttttttta tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac
     4141 gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac
     4201 atttatcaag aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa attgatacta
     4261 atggctcaac gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag
     4321 ggcaccacac aaaaagttag gtgtaacaga aaatcatgaa actatgattc ctaatttata
     4381 tattggagga ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct
     4441 gaccatttga tggagtttaa gtcaatacct tcttgaacca tttcccataa tggtgaaagt
     4501 tccctcaaga attttactct gtcagaaacg gccttaacga cgtagtcgag agtactcgta
     4561 tcgcatgtcg gtgcgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
     4621 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
     4681 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
     4741 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
     4801 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
     4861 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
     4921 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
     4981 gtagtcgacc tcctcttcag tactaaatct accaatacca aatctgatgg aagaatgggc
     5041 taatgcatca tccttaccca gcgccgagcg gccgcgtgtt acaaccaatt aaccaattct
     5101 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     5161 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     5221 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     5281 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     5341 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     5401 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     5461 tgcgcctgag cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     5521 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     5581 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     5641 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     5701 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     5761 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     5821 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     5881 ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     5941 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     6001 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     6061 gatcagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6361 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6421 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6481 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6541 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6601 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6661 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6721 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6781 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgcca
     6841 atacccagaa gaacagtaaa ataaagcaag gtacgtgaaa ttaatatttt taaatggttc
     6901 taaccgatgc cgaagaactg cgcagtccgg ttataacgtc tgacatgtcc ttttttgatt
     6961 tggaatccaa ccactcaagt gactctgttc atttactttg cgaaaaatat acccacaaat
     7021 tgcccatcga aagtgaatcg caaaccacct tcagactggc accgacaaag caaagattat
     7081 acagacagag tactttatac gtaccgttaa gtctcaagca aagggttttc ttatttactg
     7141 aacgggtaaa gagtatctgg gccggcttgc caagatgcaa accgaataag tatttcaaag
     7201 ttgcatttgc cttagccgtc ctgacaccat tggctatttg gatattttat attgactttc
     7261 gtgtacattg atcacatcga ctgttctatt ggcaaatgaa ccacgggcat tgactatttt
     7321 tcaggttact actatatatt atcatcacgg gcaaggattg taccctgaat tcgcatctag
     7381 actgatgaga cgtggtagag ccacaaacag ccggtacaag caacgatctc caggaccatc
     7441 tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa cccacagtac
     7501 agacactgcg acaacgtggc aattcgtcgc aatacaacg
//
