LOCUS       sequence-431877-        7380 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express Pex14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  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 7380)
  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: pJJB1334
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1748..1935
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1803..1827)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2411..2818
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2819..3913
                     /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(4944..5759)
                     /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(4944..5759)
                     /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     5667..5686
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5751..5770
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5984..6572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6473..6492
                     /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 gtgacgtggt
      721 cagtaaagat cgtaaggcat tgttcgactc agcagtatcc tttttaaagg atgagtccat
      781 taaagatgct ccacttttaa agaaaatcga atttttaaaa tccaaagggt taacagaaaa
      841 ggagattgaa atagccatga aagagcccaa gaaagacggt atcgtaggcg atgaagtatc
      901 gaaaaaaatt ggtagtactg agaatagagc ctcacaggat atgtatctct atgaagcgat
      961 gccaccaacg ctgccccaca gggattggaa ggactatttt gtgatggcta ctgccacagc
     1021 tgggctgttg tatggtgcat atgaagtaac tagaaggtat gtgataccaa atattttacc
     1081 agaagcaaaa agcaagttgg aaggggacaa aaaagaaatt gatgatcagt tctccaaaat
     1141 cgatacagtc ctcaatgcca tcgaagcgga gcaagctgag tttaggaaaa aggaaagcga
     1201 aacattaaag gaacttagtg acacgattgc tgaactgaaa caggcgcttg tgcagacaac
     1261 aagaagcagg gaaaagatcg aggatgagtt tagaatagtt aaactcgagg tggtcaatat
     1321 gcaaaatacg atcgacaaat ttgtttcaga taatgacggc atgcaagagt taaataatat
     1381 ccaaaaagaa atggaatctc tgaaaagctt aatgaataac cgtatggaat ccggtaatgc
     1441 gcaggacaac agattatttt ccatatctcc taatggtata cctggcatag atacgattcc
     1501 atctgcgtct gagattcttg ccaaaatggg catgcaagaa gaaagtgata aagaaaagga
     1561 aaacggcagc gatgctaata aagatgacaa tgctgttcca gcgtggaaaa aagcaagaga
     1621 acaaactatt gatagcaacg cctccattcc agaatggcaa aaaaataccg ccgccaatga
     1681 gatcagtgtc cctgactggc aaaatggaca ggtcgaggat tccatcccat agggatccta
     1741 actcgaggcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
     1801 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
     1861 taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga
     1921 ccacacctct accggcatgc cgagcaaatg cctgcaaatc gctccccatt tcgctggaaa
     1981 tctgctcgtc agtggtgctc acactgacga atcatgtaca gatcataccg atgactgcct
     2041 ggcgactcac aactaagcaa gacagccgga accagcgccg gcgaacacca ctgcatatat
     2101 ggcatatcac aacagtccac gtctcaagca gttacagaga tgttacgaac cactagtgca
     2161 ctgcagtaca tcgaggagaa cttctagtat atctacatac ctaatattat tgccttatta
     2221 aaaatggaat cccaacaatt acatcaaaat ccacattctc ttcaaaatca attgtcctgt
     2281 acttccttgt tcatgtgtgt tcaaaaacgt tatatttata ggataattat actctatttc
     2341 tcaacaagta attggttgtt tggccgagcg gtctaaggcg cctgattcaa gaaatatctt
     2401 gaccgcagtt aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta
     2461 gcaaccatta tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat
     2521 caaattcgat gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt
     2581 ttcaactgaa aaattgggag aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat
     2641 agaatagaga agcgttcatg actaaatgct tgcatcacaa tacttgaagt tgacaatatt
     2701 atttaaggac ctattgtttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt
     2761 tcttaccttt tacatttcag caatatatat atatatattt caaggatata ccattctaat
     2821 gtctgcccct aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc
     2881 cgaagccatt aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt
     2941 cgaaaatcat ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga
     3001 ggcgctggaa gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa
     3061 atggggaacc ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca
     3121 attgtacgcc aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc
     3181 aatcaagcca caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg
     3241 tatttacttt ggtaagagaa aggaagacga tggtgatggt gtcgcttggg atagtgaaca
     3301 atacaccgtt ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca
     3361 tgagccacca ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt
     3421 atggagaaaa actgtggagg aaaccatcaa gaacgaattt cctacattga aggttcaaca
     3481 tcaattgatt gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat
     3541 tataatcacc agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg
     3601 ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt
     3661 tggtttgtac gaaccatgcc acggttctgc tccagatttg ccaaagaata aggtcaaccc
     3721 tatcgccact atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga
     3781 aggtaaggcc attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga
     3841 tttaggtggt tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa
     3901 aatccttgct taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga
     3961 aattcataat agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa
     4021 ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg taaaggaata
     4081 caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac
     4141 attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga
     4201 aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa
     4261 ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc
     4321 atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg
     4381 acgtagtcga gagtactcgt atcgcatgtc ggtgcgacac gaaattacaa aatggaatat
     4441 gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa
     4501 aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga
     4561 aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag
     4621 gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa
     4681 aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa
     4741 gtcaatacct tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct
     4801 gtcagaaacg gccttaacga cgtagtcgac ctcctcttca gtactaaatc taccaatacc
     4861 aaatctgatg gaagaatggg ctaatgcatc atccttaccc agcgccgagc ggccgcgtgt
     4921 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     4981 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     5041 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     5101 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     5161 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct
     5221 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     5281 aaaccgttat tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa
     5341 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     5401 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     5461 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     5521 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     5581 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     5641 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     5701 tccatgttgg aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata
     5761 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     5821 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat
     5881 cgaacttttg ctgagttgaa ggatcagtca tgaccaaaat cccttaacgt gagttttcgt
     5941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6121 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6361 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6601 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6661 tggataaccg tgcggccgcc aatacccaga agaacagtaa aataaagcaa ggtacgtgaa
     6721 attaatattt ttaaatggtt ctaaccgatg ccgaagaact gcgcagtccg gttataacgt
     6781 ctgacatgtc cttttttgat ttggaatcca accactcaag tgactctgtt catttacttt
     6841 gcgaaaaata tacccacaaa ttgcccatcg aaagtgaatc gcaaaccacc ttcagactgg
     6901 caccgacaaa gcaaagatta tacagacaga gtactttata cgtaccgtta agtctcaagc
     6961 aaagggtttt cttatttact gaacgggtaa agagtatctg ggccggcttg ccaagatgca
     7021 aaccgaataa gtatttcaaa gttgcatttg ccttagccgt cctgacacca ttggctattt
     7081 ggatatttta tattgacttt cgtgtacatt gatcacatcg actgttctat tggcaaatga
     7141 accacgggca ttgactattt ttcaggttac tactatatat tatcatcacg ggcaaggatt
     7201 gtaccctgaa ttcgcatcta gactgatgag acgtggtaga gccacaaaca gccggtacaa
     7261 gcaacgatct ccaggaccat ctgaatcatg cgcggatgac acgaactcac gacggcgatc
     7321 acagacatta acccacagta cagacactgc gacaacgtgg caattcgtcg caatacaacg
//
