LOCUS       sequence-318568-        6125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tNCS-ePTS1 Cen6/ARS4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  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 6125)
  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: pPSG569
FEATURES             Location/Qualifiers
     source          1..6125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..705
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     575..598
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     promoter        1836..2243
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2244..3338
                     /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    3820..4323
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4367..5182)
                     /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(4367..5182)
                     /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     5090..5109
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5174..5193
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5407..5995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5896..5915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actggccgat aattgcagac gaacgcagtt cgagtttatc attatcaata ctgccatttc
       61 aaagaatacg taaataatta atagtagtga ttttcctaac tttatttagt caaaaaatta
      121 gccttttaat tctgctgtaa cccgtacatg cccaaaatag ggggcgggtt acacagaata
      181 tataacatcg taggtgtctg ggtgaacagt ttattcctgg catccactaa atataatgga
      241 gcccgctttt taagctggca tccagaaaaa aaaagaatcc cagcaccaaa atattgtttt
      301 cttcaccaac catcagttca taggtccatt ctcttagcgc aactacagag aacaggggca
      361 caaacaggca aaaaacgggc acaacctcaa tggagtgatg caacctgcct ggagtaaatg
      421 atgacacaag gcaattgacc cacgcatgta tctatctcat tttcttacac cttctattac
      481 cttctgctct ctctgatttg gaaaaagctg aaaaaaaagg ttgaaaccag ttccctgaaa
      541 ttattcccct acttgactaa taagtatata aagacggtag gtattgattg taattctgta
      601 aatctatttc ttaaacttct taaattctac ttttatagtt agtctttttt ttagttttaa
      661 aacaccaaga acttagtttc gaataaacac acataaacaa acaaaagatc tatggaagaa
      721 actgtaatgt tatatcacga attggaagtc gcagcctcag ctgacgaagt atggtctgtc
      781 gaaggttctc ctgaattagg tttacattta ccagatttgt taccagctgg aatattcgca
      841 aagtttgaga taactggaga tggtggagaa ggttctattt tggacatgac atttccacct
      901 ggtcaattcc ctcatcatta tagagagaag ttcgttttct tcgaccataa gaatagatat
      961 aagttggttg aacaaattga cggtgatttc tttgatttgg gagtaaccta ctacatggac
     1021 actattaggg tcgtcgcaac aggtcctgat tcttgtgtca tcaaatcaac aactgaatat
     1081 catgttaaac cagaatttgc taagattgtt aaacctttaa tcgatacagt tccattagcc
     1141 attatgtcag aggcaatagc taaggtagta ttggaaaata agcacaaatc ttctgaggga
     1201 tccggttcat tgggaagagg tagaagatcc aaattgtaat aactcgagtg gcagtgcttt
     1261 taactaagaa ttattagtct tttctgctta ttttttcatc atagtttaga acactttata
     1321 ttaacgaata gtttatgaat ctatttaggt ttaaaaattg atacagtttt ataagttact
     1381 ttttcaaaga ctcgtgctgt ctattgcata atgcactgga aggggaaaaa aaaggtgcac
     1441 acgcgtggct ttttcttgaa tttgcagttt gaaaaatgct gccaaaccag atgtcaacac
     1501 agctacaacg actagagatc tatgtgagga tcctaactcg agatcgagct gagcagttac
     1561 agagatgtta cgaaccacta gtgcactgca gtacatcgag gagaacttct agtatatcta
     1621 catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca
     1681 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
     1741 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
     1801 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
     1861 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
     1921 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
     1981 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
     2041 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
     2101 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
     2161 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
     2221 tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt ttgccaggtg
     2281 accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct atttctgatg
     2341 ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct gctatcgatg
     2401 ctacaggtgt tccacttcca gatgaggcgc tggaagcctc caagaaggct gatgccgttt
     2461 tgttaggtgc tgtgggtggt cctaaatggg gaaccggtag tgttagacct gaacaaggtt
     2521 tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt aactttgcat
     2581 ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt actgacttcg
     2641 ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa gacgatggtg
     2701 atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga atcacaagaa
     2761 tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc ttggataaag
     2821 ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc atcaagaacg
     2881 aatttcctac attgaaggtt caacatcaat tgattgattc tgccgccatg atcctagtta
     2941 agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt gatatcatct
     3001 ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg tccttggcct
     3061 ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt tctgctccag
     3121 atttgccaaa gaataaggtc aaccctatcg ccactatctt gtctgctgca atgatgttga
     3181 aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt aaaaaggttt
     3241 tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc gaagtcggtg
     3301 atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt tttttatgat
     3361 atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg
     3421 gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt atcaagaagg
     3481 agaaaaagga ggatgtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga
     3541 taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa
     3601 agttaggtgt aacagaaaat catgaaacta tgattcctaa tttatatatt ggaggatttt
     3661 ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc atttgatgga
     3721 gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc tcaagaattt
     3781 tactctgtca gaaacggcct taacgacgta gtcgagagta tcacgtgcta taaaaataat
     3841 tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat
     3901 taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca
     3961 aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct
     4021 tgtctgtgta gaaaaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat
     4081 cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct
     4141 ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta
     4201 ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt
     4261 aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg
     4321 atccgtcccg agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat
     4381 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     4441 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     4501 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     4561 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     4621 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     4681 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc
     4741 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     4801 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4861 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     4921 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     4981 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     5041 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     5101 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg
     5161 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     5221 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     5281 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tcatgaccaa
     5341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5521 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6061 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gcccctgaat tcgcatctag
     6121 actga
//
