LOCUS       sequence-311877-       10201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VioA, VioB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10201)
  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 10201)
  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: pJAS1052
FEATURES             Location/Qualifiers
     source          1..10201
                     /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        2474..2887
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6436..7239
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(7881..8696)
                     /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(7881..8696)
                     /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     8604..8623
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8688..8707
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8921..9509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9410..9429
                     /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 tctatgaaac attcttccga tatctgcatt gttggtgctg gtatttctgg
      721 tttgacgtgc gcaagccatc tgctggacag cccggcatgc cgtggtctga gcctgcgtat
      781 ctttgacatg cagcaagaag ccggtggccg tatccgcagc aaaatgctgg atggtaaggc
      841 aagcattgaa ctgggcgcag gtcgctactc ccctcagttg cacccgcatt tccaaagcgc
      901 aatgcagcac tatagccaaa agagcgaagt ctatccgttc acccagttga agttcaaatc
      961 tcacgtgcag caaaagctga agcgcgccat gaatgaactg tccccgcgtc tgaaagagca
     1021 tggtaaagag agctttttgc agtttgtcag ccgttatcaa ggtcacgata gcgcggttgg
     1081 tatgatccgc tctatgggtt acgacgcact gttcctgccg gatatcagcg cagaaatggc
     1141 ctacgacatt gtgggtaagc acccggagat ccagagcgtg acggacaacg acgcgaacca
     1201 atggtttgca gcggaaacgg gctttgctgg tctgattcag ggcatcaagg ctaaggttaa
     1261 ggcggcaggt gcgcgtttta gcctgggtta tcgtctgctg agcgtccgta ccgacggtga
     1321 cggctacctg ctgcaactgg caggtgacga cggctggaaa ctggagcacc gtacccgcca
     1381 tctgattctg gcgattccgc cgagcgcgat ggcgggtttg aatgttgatt ttccagaagc
     1441 ctggtccggt gcgcgctatg gcagcctgcc gctgtttaag ggctttctga cgtacggtga
     1501 gccgtggtgg ttggactaca aactggacga tcaggtgctg attgttgaca acccgctgcg
     1561 caaaatctat ttcaaaggcg ataagtacct gttcttctat accgatagcg agatggcgaa
     1621 ttactggcgc ggttgtgtcg cggagggcga ggacggttac ctggagcaaa ttcgcaccca
     1681 tttggctagc gcactgggta tcgtccgtga acgtatcccg caaccgctgg cacacgttca
     1741 caagtattgg gcgcacggcg ttgagttttg ccgtgattct gatattgacc acccgagcgc
     1801 actgtctcat cgcgacagcg gtatcatcgc gtgctccgat gcgtacacgg agcattgtgg
     1861 ttggatggag ggcggtctgc tgagcgcccg tgaggcaagc cgtctgctgt tgcagcgtat
     1921 cgccgcggga tcctaactcg agagcttttg attaagcctt ctagtccaaa aaacacgttt
     1981 ttttgtcatt tatttcattt tcttagaata gtttagttta ttcattttat agtcacgaat
     2041 gttttatgat tctatatagg gttgcaaaca agcatttttc attttatgtt aaaacaattt
     2101 caggtttacc ttttattctg cttgtggtga cgcgtgtatc cgcccgctct tttggtcacc
     2161 catgtatgct gccaaaccag atgtcaacac agctacaacg ccttgccaac agggagttct
     2221 tcagagacat ggaggctcaa aacgaaatta ttgacagcct agacatcaat agtcatacaa
     2281 cagaaagcga ccacccaact ttggctgata atagcgtata aacaatgcat actttgtacg
     2341 ttcaaaatac aatgcagtag atatatttat gcatattaca tataatacat atcacatagg
     2401 aagcaacagg cgcgttggac ttttaatttt cgaggaccgc gaatccttac atcacaccca
     2461 atcccccaca agtgatcccc cacacaccat agcttcaaaa tgtttctact ccttttttac
     2521 tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag
     2581 catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt
     2641 tggaaaagaa aaaagacacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat
     2701 ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac
     2761 ctcccattga tatttaagtt aataaacggt catcaatttc tcaagtttca gtttcatttt
     2821 tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta
     2881 atctaagttt taattacaaa agatctatga gcattctgga tttcccgcgt atccacttcc
     2941 gtggctgggc ccgtgtcaat gcgccgaccg cgaaccgcga tccgcacggc cacatcgata
     3001 tggccagcaa taccgtggcg atggcgggtg agccgttcga cctggcacgc catcctacgg
     3061 agttccaccg tcacctgcgc tccctgggtc cgcgcttcgg cttggatggt cgtgctgacc
     3121 cggaaggccc gttcagcctg gccgagggct acaacgctgc cggtaacaac cacttttcgt
     3181 gggagagcgc aaccgttagc cacgtgcaat gggatggcgg tgaggcggat cgtggtgacg
     3241 gtctggtcgg tgctcgtttg gcactgtggg gtcactacaa tgattatctg cgtaccacct
     3301 tcaatcgtgc tcgttgggtc gacagcgacc cgacgcgccg tgacgctgca caaatctatg
     3361 cgggccaatt caccattagc ccggctggtg ccggtccggg tacgccgtgg ctgtttacgg
     3421 cagacattga tgatagccat ggtgcacgtt ggacgcgtgg cggccacatt gcagagcgtg
     3481 gcggccactt cttggatgaa gagtttggtc tggcacgcct gtttcagttc tctgtgccga
     3541 aagatcaccc acattttctg tttcacccgg gtccgtttga ttccgaggcc tggcgtcgtc
     3601 tgcaattggc tctggaggat gacgacgttc tgggtctgac cgtgcaatat gcgttgttca
     3661 atatgagcac cccgcctcag ccgaacagcc cggtttttca cgatatggtc ggtgttgtcg
     3721 gtctgtggcg tcgtggtgaa ctggcgagct acccggctgg tcgtctgctg cgtccgcgtc
     3781 aaccgggtct gggtgacctg accctgcgcg tcaacggtgg tcgcgttgcg ctgaatttgg
     3841 cgtgtgccat tccgttcagc actcgtgccg cgcagccaag cgcaccggac cgcctgaccc
     3901 cggacctggg tgccaaactg ccgctgggcg atctgctgct gcgtgatgag gacggcgcac
     3961 tgttggcacg tgtgccgcag gctctgtacc aagactattg gacgaatcac ggtattgtgg
     4021 acctgccgct gctgcgcgaa ccgcgtggta gcttgaccct gagcagcgaa ctggcggagt
     4081 ggcgtgagca agactgggtc acccaaagcg acgcgtctaa cctgtacctg gaggcaccgg
     4141 atcgccgtca cggtcgcttt ttccctgaga gcatcgcgct gcgcagctac tttcgcggtg
     4201 aagcgcgtgc gcgtccggat atcccgcatc gtatcgaggg catgggcctg gtcggcgtcg
     4261 aatctcgtca ggatggcgac gctgcggaat ggcgtctgac gggtctgcgt ccgggtccgg
     4321 cacgcattgt tctggacgat ggtgccgagg cgatccctct gcgtgttctg cctgacgatt
     4381 gggcgctgga tgacgcgacc gtcgaagaag tggattacgc ctttttgtac cgccacgtta
     4441 tggcgtatta cgagctggtg tatccattca tgagcgacaa ggtgttttcc ctggctgatc
     4501 gttgcaaatg tgaaacgtac gcacgtctga tgtggcagat gtgtgatccg cagaaccgca
     4561 acaagtccta ttacatgccg agcacccgcg aactgtcggc accgaaagct cgtttgttct
     4621 tgaagtatct ggcccacgtg gaaggccagg cacgcctgca agcacctccg ccagcgggtc
     4681 cggcacgcat tgaatctaaa gcccagttgg cggcagagct gcgtaaagcc gtcgacctgg
     4741 agctgtctgt gatgctgcaa tacctgtacg cggcgtatag cattccgaac tatgcacagg
     4801 gccaacaacg tgttcgtgac ggtgcgtgga ccgccgagca gctgcaactg gcgtgcggta
     4861 gcggtgaccg tcgccgtgat ggcggtattc gtgcagcact gctggaaatt gctcatgaag
     4921 aaatgattca ttacctggtc gttaacaacc tgctgatggc cctgggcgag ccgttctacg
     4981 cgggtgtccc gctgatgggc gaagcggcac gtcaggcgtt tggcctggac accgagttcg
     5041 ctctggaacc gtttagcgaa agcacgctgg cacgttttgt tcgtctggaa tggccgcact
     5101 ttatcccagc accgggcaaa tccatcgcgg actgctatgc cgccattcgt caggcgtttt
     5161 tggatctgcc ggacttgttt ggtggcgagg caggtaagcg tggcggtgaa caccacctgt
     5221 tcctgaatga gctgaccaac cgtgcgcatc cgggttatca actggaagtt ttcgatcgcg
     5281 actcggcgct gtttggtatt gcatttgtga ccgatcaggg cgaaggtggc gctctggaca
     5341 gcccgcacta cgaacatagc cattttcaac gtctgcgtga aatgagcgcg cgtatcatgg
     5401 ctcaaagcgc accgttcgaa ccggcgctgc cggcgttgcg taatccggtt ctggatgaga
     5461 gcccgggttg ccaacgtgtc gcagacggtc gtgcgcgtgc gctgatggca ttgtaccaag
     5521 gcgtttatga gctgatgttt gcgatgatgg cgcagcactt cgccgtgaaa ccgctgggta
     5581 gcttgcgtcg cagccgcctg atgaacgcag caatcgatct gatgaccggt ctgttgcgtc
     5641 cgctgagctg cgcgctgatg aacctgccaa gcggcatcgc cggtcgcacg gccggtccgc
     5701 cgctgccggg tccggttgac acccgtagct atgacgacta cgcgctgggc tgtcgcatgc
     5761 tggcacgccg ttgcgagcgt ctgctggagc aggcgagcat gctggaaccg ggttggctgc
     5821 cggatgcgca gatggagctg ctggatttct atcgtcgcca aatgctggac ttggcgtgcg
     5881 gcaaactgag ccgcgaggcc ggatcctaac tcgagattga attgaattga aatcgataga
     5941 tcaatttttt tcttttctct ttccccatcc tttacgctaa aataatagtt tattttattt
     6001 tttgaatatt ttttatttat atacgtatat atagactatt atttatcttt taatgattat
     6061 taagattttt attaaaaaaa aattcgctcc tcttttaatg cctttatgca gttttttttt
     6121 cccattcgat atttctatgt gctggatgca cacactggct taagatgaca acgactagag
     6181 atctatgtga ggatcctaac tcgagatcga gctgagcagt tacagagatg ttacgaacca
     6241 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
     6301 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     6361 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
     6421 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     6481 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     6541 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     6601 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     6661 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     6721 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     6781 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     6841 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     6901 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     6961 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     7021 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
     7081 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
     7141 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     7201 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     7261 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
     7321 tagagcactt gaatccactg ccccgggaat ctcggtcgta atgatttcta taatgacgaa
     7381 aaaaaaaaaa ttggaaagaa aaagcttcat ggcctttata aaaaggaact atccaatacc
     7441 tcgccagaac caagtaacag tattttacgg ggcacaaatc aagaacaata agacaggact
     7501 gtaaagatgg acgcattgaa ctccaaagaa caacaagagt tccaaaaagt agtggaacaa
     7561 aagcaaatga aggatttcat gcgtttgtac tctaatctgg tagaaagatg tttcacagac
     7621 tgtgtcaatg acttcacaac atcaaagcta accaataagg aacaaacatg catcatgaag
     7681 tgctcagaaa agttcttgaa gcatagcgaa cgtgtagggc agcgtttcca agaacaaaac
     7741 gctgccttgg gacaaggctt gggccgataa ggtgtactgg cgtatatata tctaattatg
     7801 tatctctggt gtagcccatt tttagcatgt aaatataaag accgagcggc cgcgtgttac
     7861 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     7921 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     7981 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     8041 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     8101 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     8161 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     8221 ccgttattca ttcgtgattg cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga
     8281 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     8341 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     8401 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     8461 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     8521 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     8581 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     8641 atgttggaat ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca
     8701 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     8761 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     8821 acttttgctg agttgaagga tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8881 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     9001 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     9061 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     9121 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     9181 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     9241 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     9301 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9361 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9421 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9481 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     9541 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     9601 ataaccgtgc ggccgccaat gctaaattcg agtgaaacac aggaagatca gaaaatcctc
     9661 atttcatcca tattaacaat aatttcaaat gtttatttgc attatttgaa actaggcaag
     9721 acaagcaacg aaacgttttt gaaaattttg agtattttca ataaatttgt agaggactca
     9781 gatattgaaa aaaagctaca gcaattaata cttgataaga agagtattga gaagggcaac
     9841 ggttcatcat ctcatggatc tgcacatgaa caaacaccag agtcaaacga cgttgaaatt
     9901 gaggctactg cgccaattga tgacaataca gacgatgata acaaaccgaa gttatctgat
     9961 gtagaaaagg attaaagatg ctaagagata gtgatgatat ttcataaata atgtaattct
    10021 atatatgtta attacctttt ttgcgaggca tatttatggt gaaggataag ttttgaccat
    10081 caaagaaggt taatgtggct gtggtttcag ggtccataaa gcccacatgg ataacattac
    10141 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgca gttcgagttt
    10201 a
//
