LOCUS       sequence-311871-        6421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VioE-dead_ePTS1  CEN6/ARS4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6421)
  AUTHORS   Grewal PS, Samson JA, Baker JJ, Choi B, Dueber JE
  TITLE     Peroxisome compartmentalization of a toxic enzyme improves alkaloid 
            production.
  JOURNAL   Nat Chem Biol. 2020 Oct 12. pii: 10.1038/s41589-020-00668-4. doi: 
            10.1038/s41589-020-00668-4.
  PUBMED    33046851
REFERENCE   2  (bases 1 to 6421)
  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. 2020 Oct 12. pii: 10.1038/s41589-020-00668-4. doi: 
            10.1038/s41589-020-00668-4."
COMMENT     Sequence Label: pJAS1669
FEATURES             Location/Qualifiers
     source          1..6421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..667
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     537..560
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      1302..1489
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1357..1381)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1965..2372
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2373..3467
                     /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"
     misc_feature    3949..4452
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4496..5311)
                     /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(4496..5311)
                     /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     5219..5238
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5303..5322
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5536..6124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6025..6044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
       61 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
      121 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
      181 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
      241 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
      301 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
      361 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
      421 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
      481 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
      541 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      601 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac
      661 aaacaaaaga tctatggaga accgtgagcc accactgttg ccagcccgtt ggagcagcgc
      721 ctatgtctct tattggagcc cgatgctgcc ggatgaccag ctgaccagcg gctattgctg
      781 gttcgactat gaacgtgaca tctgtcgtat tgacggcctg ttcaatccgt ggagcgagcg
      841 tgatactggt tatcgcctgt ggatgtcgga ggttggtaat gcggccagcg gccgtacctg
      901 gaaacaaaaa gtcgcctatg gtcgtgagcg taccgccctg ggtgaacagc tgtgtgagcg
      961 tccgctggat gatgagactg gcccttttgc cgaattgttc ctgccacgcg atgtcctgcg
     1021 ccgtctgggt gcccgtcaca ttggccgtcg cgtggttctg ggtcgcgaag cggacggttg
     1081 gcgttaccag cgcccaggta aaggtccgag caccctgtac ctggatgcgg cgagcggcac
     1141 tccactgcgc atggtcaccg gcgatgaagc gtcgcgtgca agcctgcgtg attttccgaa
     1201 tgtgagcgag gcggagatcc cggacgcggt tttcgcggcc aagcgcggat ccggttcatt
     1261 gggaagaggt agaagatcca aaacctaata actcgagtgg cgcgaatttc ttatgattta
     1321 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     1381 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     1441 tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccgagca
     1501 aatgcctgca aatcgctccc catttcgctg gaaatctgct cgtcagtggt gctcacactg
     1561 acgaatcatg tacagatcat accgatgact gcctggcgac tcacaactaa gcaagacagc
     1621 cggaaccagc gccggcgaac accactgcat atatggcata tcacaacagt ccacgtctca
     1681 agcagttaca gagatgttac gaaccactag tgcactgcag tacatcgagg agaacttcta
     1741 gtatatctac atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca
     1801 aaatccacat tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa
     1861 acgttatatt tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg
     1921 agcggtctaa ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc
     1981 aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat
     2041 aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg
     2101 ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag
     2161 gaaaggtgag agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
     2221 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
     2281 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
     2341 atatatatat atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt
     2401 tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta
     2461 tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg
     2521 ctatcgatgc tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg
     2581 atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg aaccggtagt gttagacctg
     2641 aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta
     2701 actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta
     2761 ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag
     2821 acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa
     2881 tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct
     2941 tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca
     3001 tcaagaacga atttcctaca ttgaaggttc aacatcaatt gattgattct gccgccatga
     3061 tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg
     3121 atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt
     3181 ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt
     3241 ctgctccaga tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa
     3301 tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta
     3361 aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg
     3421 aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt
     3481 ttttatgata tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat
     3541 tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta catacattta
     3601 tcaagaagga gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc
     3661 tcaacgtgat aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac
     3721 cacacaaaaa gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg
     3781 gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca
     3841 tttgatggag tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct
     3901 caagaatttt actctgtcag aaacggcctt aacgacgtag tcgagagtat cacgtgctat
     3961 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     4021 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     4081 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     4141 gtttcatctt gtctgtgtag aaaaccacac acgaaaatcc tgtgatttta cattttactt
     4201 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     4261 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     4321 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     4381 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     4441 aggcaagcga tccgtcccga gcggccgcgt gttacaacca attaaccaat tctgattaga
     4501 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     4561 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     4621 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     4681 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     4741 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     4801 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     4861 gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     4921 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     4981 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     5041 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     5101 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     5161 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     5221 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg
     5281 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     5341 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     5401 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagt
     5461 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5521 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5581 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5641 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5701 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5761 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5821 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5881 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5941 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6001 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6061 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6121 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6181 catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg cccctgaatt
     6241 cgcatctaga ctgatgagac gtggtagagc cacaaacagc cggtacaagc aacgatctcc
     6301 aggaccatct gaatcatgcg cggatgacac gaactcacga cggcgatcac agacattaac
     6361 ccacagtaca gacactgcga caacgtggca attcgtcgca atacaacgca gttcgagttt
     6421 a
//
