LOCUS       sequence-311858-        7045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tNCS-Venus-ePTS1 CEN6/ARS4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7045)
  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 7045)
  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: pPSG1066
FEATURES             Location/Qualifiers
     source          1..7045
                     /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"
     CDS             1172..1882
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     promoter        2589..2996
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2997..4091
                     /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    4573..5076
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(5120..5935)
                     /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(5120..5935)
                     /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     5843..5862
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5927..5946
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6160..6748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6649..6668
                     /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 tctatggaag aaactgtaat gttatatcac gaattggaag tcgcagcctc
      721 agctgacgaa gtatggtctg tcgaaggttc tcctgaatta ggtttacatt taccagattt
      781 gttaccagct ggaatattcg caaagtttga gataactgga gatggtggag aaggttctat
      841 tttggacatg acatttccac ctggtcaatt ccctcatcat tatagagaga agttcgtttt
      901 cttcgaccat aagaatagat ataagttggt tgaacaaatt gacggtgatt tctttgattt
      961 gggagtaacc tactacatgg acactattag ggtcgtcgca acaggtcctg attcttgtgt
     1021 catcaaatca acaactgaat atcatgttaa accagaattt gctaagattg ttaaaccttt
     1081 aatcgataca gttccattag ccattatgtc agaggcaata gctaaggtag tattggaaaa
     1141 taagcacaaa tcttctgagg gatccgggtc ctctaaaggt gaagaattat tcactggtgt
     1201 tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg
     1261 tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaattgattt gtactactgg
     1321 taaattgcca gttccatggc caaccttagt cactacttta ggttatggtt tgcaatgttt
     1381 tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg
     1441 ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga
     1501 agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa
     1561 agaaggtggt aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta
     1621 catcactgct gacaaacaaa agaatggtat caaagctaac ttcaaaatta gacacaacat
     1681 tgaagatggt ggtgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg
     1741 tccagtcttg ttaccagaca accattactt atcctatcaa tctgccttat ccaaagatcc
     1801 aaacgaaaag agagatcaca tggtcttgtt agaatttgtt actgctgctg gtattaccca
     1861 tggtatggat gaattgtaca aagggtcctt gggaagaggt agaagatcca aattgtaata
     1921 atggcagtgc ttttaactaa gaattattag tcttttctgc ttattttttc atcatagttt
     1981 agaacacttt atattaacga atagtttatg aatctattta ggtttaaaaa ttgatacagt
     2041 tttataagtt actttttcaa agactcgtgc tgtctattgc ataatgcact ggaaggggaa
     2101 aaaaaaggtg cacacgcgtg gctttttctt gaatttgcag tttgaaaaat gctggaaatc
     2161 tgctcgtcag tggtgctcac actgacgaat catgtacaga tcataccgat gactgcctgg
     2221 cgactcacaa ctaagcaaga cagccggaac cagcgccggc gaacaccact gcatatatgg
     2281 catatcacaa cagtccacgt ctcaagcagt tacagagatg ttacgaacca ctagtgcact
     2341 gcagtacatc gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa
     2401 aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac
     2461 ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc
     2521 aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga
     2581 ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc
     2641 aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca
     2701 aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt
     2761 caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag
     2821 aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat
     2881 ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc
     2941 ttacctttta catttcagca atatatatat atatatttca aggatatacc attctaatgt
     3001 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
     3061 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
     3121 aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg
     3181 cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat
     3241 ggggaaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
     3301 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
     3361 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
     3421 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
     3481 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
     3541 agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat
     3601 ggagaaaaac tgtggaggaa accatcaaga acgaatttcc tacattgaag gttcaacatc
     3661 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
     3721 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
     3781 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
     3841 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta
     3901 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
     3961 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
     4021 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
     4081 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
     4141 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
     4201 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
     4261 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
     4321 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
     4381 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
     4441 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
     4501 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
     4561 gtagtcgaga gtatcacgtg ctataaaaat aattataatt taaatttttt aatataaata
     4621 tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg
     4681 aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct
     4741 gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaaaacc acacacgaaa
     4801 atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt
     4861 tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt
     4921 ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
     4981 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
     5041 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc ccgagcggcc gcgtgttaca
     5101 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     5161 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     5221 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     5281 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     5341 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
     5401 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     5461 cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac
     5521 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     5581 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     5641 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     5701 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     5761 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     5821 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     5881 tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg ctcataacac
     5941 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     6001 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     6061 cttttgctga gttgaaggat cagtcatgac caaaatccct taacgtgagt tttcgttcca
     6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6841 taaccgtgcg gccgcccctg aattcgcatc tagactgatg agacgtggta gagccacaaa
     6901 cagccggtac aagcaacgat ctccaggacc atctgaatca tgcgcggatg acacgaactc
     6961 acgacggcga tcacagacat taacccacag tacagacact gcgacaacgt ggcaattcgt
     7021 cgcaatacaa cgcagttcga gttta
//
