LOCUS       sequence-311863-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tNCS-VioE-dead_ePTS1  CEN6/ARS4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  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 6704)
  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: pJAS1644
FEATURES             Location/Qualifiers
     source          1..6704
                     /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"
     promoter        2307..2714
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2715..3809
                     /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    4291..4794
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4838..5653)
                     /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(4838..5653)
                     /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     5561..5580
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5645..5664
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5878..6466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6367..6386
                     /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 gatccgagaa ccgtgagcca ccactgttgc cagcccgttg
     1201 gagcagcgcc tatgtctctt attggagccc gatgctgccg gatgaccagc tgaccagcgg
     1261 ctattgctgg ttcgactatg aacgtgacat ctgtcgtatt gacggcctgt tcaatccgtg
     1321 gagcgagcgt gatactggtt atcgcctgtg gatgtcggag gttggtaatg cggccagcgg
     1381 ccgtacctgg aaacaaaaag tcgcctatgg tcgtgagcgt accgccctgg gtgaacagct
     1441 gtgtgagcgt ccgctggatg atgagactgg cccttttgcc gaattgttcc tgccacgcga
     1501 tgtcctgcgc cgtctgggtg cccgtcacat tggccgtcgc gtggttctgg gtcgcgaagc
     1561 ggacggttgg cgttaccagc gcccaggtaa aggtccgagc accctgtacc tggatgcggc
     1621 gagcggcact ccactgcgca tggtcaccgg cgatgaagcg tcgcgtgcaa gcctgcgtga
     1681 ttttccgaat gtgagcgagg cggagatccc ggacgcggtt ttcgcggcca agcgcggaag
     1741 cggttcattg ggaagaggta gaagatccaa aacctaataa ctcgagtaat ggcagtgctt
     1801 ttaactaaga attattagtc ttttctgctt attttttcat catagtttag aacactttat
     1861 attaacgaat agtttatgaa tctatttagg tttaaaaatt gatacagttt tataagttac
     1921 tttttcaaag actcgtgctg tctattgcat aatgcactgg aaggggaaaa aaaaggtgca
     1981 cacgcgtggc tttttcttga atttgcagtt tgaaaaatgc tgagcagtta cagagatgtt
     2041 acgaaccact agtgcactgc agtacatcga ggagaacttc tagtatatct acatacctaa
     2101 tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac attctcttca
     2161 aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata tttataggat
     2221 aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct aaggcgcctg
     2281 attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg taagatgcaa
     2341 gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg
     2401 cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc
     2461 tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg
     2521 aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact
     2581 tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt
     2641 cttactttct aacttttctt accttttaca tttcagcaat atatatatat atatttcaag
     2701 gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg
     2761 gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca
     2821 atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg
     2881 ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt ttgttaggtg
     2941 ctgtgggtgg tcctaaatgg ggaaccggta gtgttagacc tgaacaaggt ttactaaaaa
     3001 tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca tccgactctc
     3061 ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca
     3121 gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg
     3181 cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga atggccgctt
     3241 tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatgttt
     3301 tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac gaatttccta
     3361 cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa
     3421 cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag
     3481 cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag
     3541 acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa
     3601 agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat
     3661 tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag
     3721 gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg
     3781 ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca
     3841 taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt
     3901 catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg
     3961 aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg ataaggaaaa
     4021 agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa aagttaggtg
     4081 taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt tctctaaaaa
     4141 aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc
     4201 aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc
     4261 agaaacggcc ttaacgacgt agtcgagagt atcacgtgct ataaaaataa ttataattta
     4321 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     4381 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     4441 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     4501 agaaaaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     4561 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
     4621 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
     4681 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
     4741 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtccc
     4801 gagcggccgc gtgttacaac caattaacca attctgatta gaaaaactca tcgagcatca
     4861 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     4921 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     4981 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     5041 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     5101 gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     5161 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagg cgaaatacgc
     5221 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     5281 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     5341 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     5401 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     5461 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     5521 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     5581 catataaatc agcatccatg ttggaattta atcgcggcct ggagcaagac gtttcccgtt
     5641 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     5701 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
     5761 tttgttgaat aaatcgaact tttgctgagt tgaaggatca gtcatgacca aaatccctta
     5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6001 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6241 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6481 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6541 atcccctgat tctgtggata accgtgcggc cgcccctgaa ttcgcatcta gactgaactg
     6601 gccgataatt gcagacgaac gactagagat ctatgtgagg atcctaactc gagatcgagc
     6661 tggttctgta tgggcacaga caacctaaac gcagttcgag ttta
//
