LOCUS       sequence-224231-       11559 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of a fungal biosynthetic gene cluster.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 11559)
  AUTHORS   Harvey CJB, Tang M, Schlecht U, Horecka J, Fischer CR, Lin HC, Li J,
            Naughton B, Cherry J, Miranda M, Li YF, Chu AM, Hennessy JR, Vandova
            GA, Inglis D, Aiyar RS, Steinmetz LM, Davis RW, Medema MH, Sattely 
            E, Khosla C, St Onge RP, Tang Y, Hillenmeyer ME
  TITLE     HEx: A heterologous expression platform for the discovery of fungal 
            natural products.
  JOURNAL   Sci Adv. 2018 Apr 11;4(4):eaar5459. doi: 10.1126/sciadv.aar5459. 
            eCollection 2018 Apr.
  PUBMED    29651464
REFERENCE   2  (bases 1 to 11559)
  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: "Sci Adv."; 
            date: "2018-04-11"; volume: "4(4):eaar5459. doi"; pages: " 
            10.1126/sciadv.aar5459. eCollection 2018 Apr"
COMMENT     Sequence Label: pCHKU36.2-2.2
FEATURES             Location/Qualifiers
     source          1..11559
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      1523..1822
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     promoter        5661..5881
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5882..6685
                     /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             8411..8422
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(11535..11551)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11535..11551)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 tatctaaaaa ttgccttatg atccgtctct ccggttacag cctgtgtaac tgattaatcc
       61 tgcctttcta atcaccattc taatgtttta attaagggat tttgtcttca ttaacggctt
      121 tcgctcataa aaatgttatg acgttttgcc cgcaggcggg aaaccatcca cttcacgaga
      181 ctgatctcct ctgccggaac accgggcatc tccaacttat aagttggaga aataagagaa
      241 tttcagattg agagaatgaa aaaaaaaaaa aaaaaaaagg tagagagcat aaaaatgggg
      301 ttcacttttt ggcaaagtta cagtatgctt attacatata aatagagtgc cgataatggc
      361 tttttttcat cttcgaaata cgcttgctac tgctcttcca gcgtttttat tacttctttc
      421 ttgtttctcc ttagtatata aaatatcaag ctacaaaaag catacaatca actatcaact
      481 attaactata tcgtaataca caatggatac ttttactatg aaaaaggaca aaggtggtta
      541 caggcagatt aataaatcgt tgaatatctg tgccttcgat gagtaccttc ggggccaaac
      601 tgcgcgacta ccgcatatag agagtgttga acagctcact ccgagagtac ttcgcattct
      661 cggtcagaac ccgggaaagt tcacgcatca aggaactaat acatacatcg tcggcaccgg
      721 ccaacgccga ctcattatcg acacaggtgg cggccagcct gaatggaccg aactaatcgc
      781 gtcgactttt aaatcaatga acataagctt atctcatgtt ctgttgaccc attggcacgg
      841 tgaccacact ggtggtgtct caggtttatt gcgcctgtat ccgtatctca cggcttcgat
      901 ctacaagaac gagccgaatc agggccagca gaacatcgct gacggacaga tattctccgt
      961 tgatggcgca acaatccatg ctatccatgc acccggacat tcagaggacc atatgtgttt
     1021 tatcctagaa gaggagaatg cgatgtttac cggcgataat atccttggcc atggtactag
     1081 tgctgtagaa gacctaggca ccttcatggc cagcctcgag aagatggccg cccagaattg
     1141 tcaaatcggt tattctgctc atggcgctac aatcgcaaac ttgccagcaa agattgctgg
     1201 agaattagaa agcaagtggc gacgagagaa gcaagttatg aaaggactgg cgcgagttcg
     1261 catccgtggc gagaaaagcg ccaccgtcag ggacctcgta actgagatct acggcggttc
     1321 gctagacaaa gacacgagga cgctcgcatt ggaaccattt attgatgaag tgttacggaa
     1381 gttggcggat gactccaagg ttgcttttga aatgagagga ggcaagaaga agtggttctc
     1441 tgtggaggag gtggaacaaa caaccacctg gcggttcaca gccacgaata aaagttcacc
     1501 agctgtacct gttcaagttt gagcggatct cttatgtctt tacgatttat agttttcatt
     1561 atcaagtatg cctatattag tatatagcat ctttagatga cagtgttcga agtttcacga
     1621 ataaaagata atattctact ttttgctccc accgcgtttg ctagcacgag tgaacaccat
     1681 ccctcgcctg tgagttgtac ccattcctct aaactgtaga catggtagct tcagcagtgt
     1741 tcgttatgta cggcatcctc caacaaacag tcggttatag tttgtcctgc tcctctgaat
     1801 cgtctccctc gatatttctc attttccttc gcatgccagc attgaaatga tcgaagttca
     1861 atgatgaaac ggtaattctt ctgtcattta ctcatctcat ctcatcaagt tatataattc
     1921 tatacggatg taatttttca cttttcgtct tgacgtccac cctataattt caattattga
     1981 accctcacga tccagttctc cagtgacaca gcctttatct ggtcaaacct ttctttctaa
     2041 tcacctatgc tgatgcttaa ttaagggatt tttgtctcca tcaacggcat gcgcccaaaa
     2101 atgacgtttt ttttaaccca tagacacgaa actacccatt ttccaccggc ctgacctacc
     2161 accggaacaa cggccatctc caacttgcaa gttggggaaa ttaagagcat cgcaggttta
     2221 atggaagaaa aaaaaaaggt acagcacagc gcaaatggag ttagttccct tatgtcacac
     2281 actcacacac agtcggtcag atcaagcata ctgggtgcgt ataaatagag tggccattgc
     2341 caccctgttt atctcaaaat ctgtcttgtt agtggtcttc tccctttttc aggttacaat
     2401 tctcttgttt ctacttagta tataagtata tcaagctata ttaagcatac tatcaactgt
     2461 caactctatc ctcaaaatac aatacaaaat ggacgaggtg agatatcgcg accagctcgc
     2521 cgcagccgtc aagagcctca tcgatagctg tcgaggagct gattcttccg accaactggc
     2581 ggcaagcgct gatactgaag aaaacaagga ctatgcttct ttaaacagag ccaggtcaag
     2641 tattcttgcc agtactgcag caatcaagtc actagttgct gaaccaaaaa acttgcttca
     2701 ggatttcgct cgtcagattg agatccttgc cagtcttcgg tggctagcag aatttcaggt
     2761 tttggcgtgc atcccgtttg acgaaagttt gcctgtcgag gacctggcag acttggtagg
     2821 agttcccgag agccagctgg tccgggttat ccgtctgacg gccacatgcg gctttttacg
     2881 agagccaata tcgggttttg tatcccacag cgctctctcg gcacaattca taaccaatca
     2941 gtctttactt gacgccactg tgtttattgc cgacttggcc actcctacag cattgcagat
     3001 gccacaagca acgcagcgct ttgggaattc ctcgagtgcc acagatagcg catacaattt
     3061 ggctttgaat acggtgcgac cgtttcatgt ggctgtccag gagcgctctc ggctgcgccg
     3121 ccaatggtcg gcctaccttt gccacgcagc tgggttgcat caagaagaag agatagttga
     3181 cgttctatcg aagctcaact ggtcaaacct tggaaatgca tgtatagttg aggtcggcgc
     3241 aaagtcaact tcaatggcgg gttctctttc caaaatgttc ccaagcttgc gattcgtcgt
     3301 gcaaattgac aacacaagat cttatttgtt agatcaggac tacgcgtggc agagtgcagt
     3361 tctaggaggc tcgacccaag atagctctgg ctccgggtct agtccttccc ccacaaaccc
     3421 tttcgtcacg gtttcttatc gcgccatggg tatgcctcag tccgtgacag atgctgccgt
     3481 atatataatc cacattcctg tggcatcatc cggattgaca attaaagcag agctggatga
     3541 ttatttgggt gttcttcgag ccagcggagg tatcatgctc atcatgacgg cacggttgct
     3601 tcccgagcca ggctcactct ccaactcaga ggctgagagt atggcgcgcg cacgggatct
     3661 gagcatgcta catctagcta acgagggtga gatggaaatt tccgatctcc tgagtactat
     3721 aaagacggtg ggggacagcg ctgggaaact cgttgttact aatcacctac gctcaaacga
     3781 cggcataatt ctagctttaa ccatcaagta tcaagcttac attattcata agcctacatt
     3841 cgatagtaga tcaccagcat ttacctccga cacactcccg atcgttggct cctggaacta
     3901 ttttacccag cacaaaaatt tttggaagga ctgcgtcgca gagtctaaga acggccactt
     3961 tagtttttgg atcggcaaga accatatcat tggcctatct ggcgcggctg ctcgcaagat
     4021 gtatctagac agccgcgatc tcgatcctat tggagggtct atgcttattg ggcatggacc
     4081 agacattata gatggtaaat catccattat acctgccatc tggacgacta ccgtaaccgg
     4141 cgaccgtctc tttgcccatc gccggcttct cgatttgcaa aaatcggaac agctcacgaa
     4201 gcgcctcgct cgtgtgacca aggactgccg cgacacattt agttcaatgt ctacaaactc
     4261 gtttggggtc ataaatccaa ctaagtcgtg ctatcccatt gttgtgaagc agtactctcg
     4321 cattgtgggc gccgatgaaa tctcgaataa cccaaagatg ctagaaaagc tccaatatta
     4381 catgcatatc ctacagacca cttcaagtgt ccatctcctt gcttttcctt ggctgtcata
     4441 ctttagtgtg gcctattgga aacgtcgcta tggccgttgg gggttttcta acattgtgac
     4501 gcctattgtc aacaagcgaa tgatgaaggg cgcacttcgc gttgatgatg cgctgcaact
     4561 tcttattgat aatggcgaca gcaaaaccta tattatcgac ttcctcatta gcatgttatt
     4621 tatcgccggt gccaacgcca ctgttttaac cggtgccatg ctaaatatca tcgcccacca
     4681 cccagactgg caagagagaa tataccgtga gattaaggct accgctgcat cccactcaac
     4741 gaacaaaggt gcatcattgg tagaccaact cgactcaatg ccattgtccg cctgggaatc
     4801 actttctcag tctcttgaac tttgtttcaa ggaaaccatc cgcatgtggg ctgttttccc
     4861 agtaagtcgc agaaatatca ccgatcatat tataaatatc gcaggaacac acgaggttat
     4921 cccacctggt ggctatactg catatcatag cactgatgta cattataacg aaaacctcta
     4981 cccggatccg atgagatggg acccggaacg cttttgcgat ggtcgcgaag agtttaaaaa
     5041 ggaggattat ggatatattg gttggggtca gggacgtcat ccttgcattg gcatgcgttg
     5101 ggctaaactc cagcaaaata taaatatcgc ctatcctcta gctatgtaca agtggagcgg
     5161 ctgcgatgag aatggacagc ccaaccctaa ctttgcacat aagccaacag taaacagagg
     5221 cgcccccact ctgccgatgg gactttactg taaatatgtt ccgcgaggag atacataaac
     5281 aaatcgctct taaatatata cctaaagaac attaaagcta tattataagc aaagatacgt
     5341 aaattttgct tatattatta tacacatatc atatttctat atttttaaga tttggttata
     5401 taatgtacgt aatgcaaagg aaataaattt tatacattat tgaacagcgt ccaagtaact
     5461 acattatgtg cactaatagt ttagcgtcgt gaagacttta ttgtgtcgcg aaaagtaaaa
     5521 attttaaaaa ttagagcacc ttgaacttgc gaaaaaggtt ctcatcaact gtttaaaagg
     5581 aggatatcag gtcctatttc tgacaaacaa tatacaaatt tagtttcaaa gatgaatcag
     5641 tgcgcgaagg acataactca ttcaattcaa ttcatcattt tttttttatt cttttttttg
     5701 atttcggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag
     5761 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa
     5821 ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat
     5881 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa
     5941 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
     6001 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
     6061 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
     6121 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     6181 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     6241 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     6301 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     6361 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     6421 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     6481 gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt
     6541 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     6601 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     6661 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     6721 ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg tgtgaaatac
     6781 cgcacagatg cgtaaggaga aaataccgca tcaggatagg aaaaaaccga gcttcctttc
     6841 atccggcgcg gctgtgttct acatatcact gaagctccgg gtattttaag ttatacaagg
     6901 gaaagatgcc ggctagacta gcaagtttta ggctgcttaa cattatggat aggcggataa
     6961 agggcccaaa caggattgta aagcttagac gcttctggtt ggacaatggt acgtttgtgt
     7021 attaagtaag gcttggctgg ggatagcaac attgggcaga gtatagaaga ccacaaaaaa
     7081 aaggtatata agggcagaga agtctttgta atgtgtgtaa cttctcttcc atgtgtaatc
     7141 agtatttcta cttacttctt aaatatacag aagtaagaca gataaccaac agcctttccc
     7201 agatatacat atatatcttt atttcagctt aaacaataat tatatttgtt taactcaaaa
     7261 ataaaaaaaa aaaccaaact cacgcaacta attattccat aataaaataa caacatgaat
     7321 accaccaaca ttgcccgtgg ggcaaatgga cacgtcaaaa taagctcgaa atgggacgag
     7381 attacagctg tcgaccgaga aacgtcagaa aagctacgat tggggcttct accctcgtcg
     7441 aagaacatag acatcaatat tctaaactct cctggtattg tatatcctag tgtatgctct
     7501 tcttccgctg tatgtcaagt aaagactgtc actacaccct ctgcagaaga catagcggtt
     7561 tcagaagcaa tagatgcaat tgaagatctt acaagcgact ttgaagctgt cacagatgag
     7621 agcaataccg aggaagttcc aagtgagctg actccaaagg tcaccaaacg gaaaaataca
     7681 cgagtttcca agctaaaata tgccgcggaa ccttctcaaa agacccaatg gccccgaacc
     7741 tggttaggta acgacagcct gtgcaacgac aagtctagtc ccaagaccat cgaatacggc
     7801 tgcaagaata gtcccgcatc atctctgtgg acagatagcg aatcttggga tactccaagc
     7861 ttcctcagat atgatgcaga aaggccctca cggtatcgat ttgtgcggcc gcgcgaaaca
     7921 gttcgccagc gagttacttc actgaagcat aaagttgttg aaattgcgag aactgatctt
     7981 ggctgggtga aagaggccat gggcaacatt cgtaccaaag ctagcccgga aaatcccaac
     8041 catagagttg gaagtgccga aaactcacgc gagagttcaa gagacgactc tacttatttg
     8101 agtctgaggg gcggagcggg cgatatttca agcttgaacg gagcgggaaa tgacattttt
     8161 gggattggcg gggggactgg tgattgttgt gctcaacttt ctgcagagtt gggatcctct
     8221 gtgggttttg aattgagcag catctatgcc aactcgcaga gttcgtattg ggcaaaacaa
     8281 caatcggagc tcactccaag ctgtattatt gtgccttcga cggcgtctga agtcgcggtt
     8341 gccatctcca tcatttcaac aatcgagtcg tgcaacttcg cagtcaaagg tgcagggcac
     8401 ggaacagttg tgggagctac aaatatcaat ggaggtgtca agtttgatat gagccgcctg
     8461 aacgaaattg agaccaactc ggaaggaact gtcaccagga tcggggccgg aagccaatgg
     8521 ggagaggtct atgagtatct tgacaacaga ggcctttccg ttgctggagg tcgaaacggc
     8581 gatgttggcg tagctggagt gctccttgga ggcggaatat cattctatgg tccccgagta
     8641 ggctgggcaa cggataatat tttgaatgtc gaagtcgtgc ttgcctcggg agagattacc
     8701 aacgccaatt ccacttcaaa ccaagaccta cttaaagcat tgaaaggcgg gaacagcaac
     8761 ttcggtatcg ccacattatt tgatctgcgc acattccccc aaggtgaaat gtgggtgggt
     8821 tatctcggtc aatccattga tagccgcgat gaagttttta aagccatcgc cgatatcgcg
     8881 tcaaatgcca atgaggaccc tttcgctgct ttggtgggcg acttcaagtt taattccgcc
     8941 accaaatctt gggtcatgaa ccatactgtg gcatatacaa agccagttgc caacccgccg
     9001 atatttcagc cattggttaa catcaagccc cagttgagca acaacattgc aatcacaaac
     9061 attagcacca ttgctacggg gcagaagaat ggagccaatg ttgcttataa caagaaccat
     9121 ctgtcttata cgggcacgta cggaaatgat gcggcgcttc tctccaagat tttcgacatc
     9181 agcaacgcca ccgttcacaa gataatgtcg cagaccaatg gtgggatcaa atggacaaca
     9241 atgatggaac catttccagc gctggtggac agctttggag ataaaaacgg cggaaactca
     9301 ctcggtttga gcgcagagac tggagatcgt attgtcatct tgcaattggc acaatttgag
     9361 gacagagatg cgaacgacat ggtcgacaaa gagcttcaca acatgtttga ggagattgca
     9421 aaagttgcat cggaaatggg actcctccgc cgctttcggt acttgaatta tgcggacaag
     9481 cagcagaacc ccattgcctc atacgggccg gaaaatgtgg cgcaattgaa ggctacaagc
     9541 aagaaatacg atccgaaggg tttattccag agacaagcgc ctggtgggtt taagcttgga
     9601 atgcagcagt gaagtgcttt taactaagaa ttattagtct tttctgctta ttttttcatc
     9661 atagtttaga acactttata ttaacgaata gtttatgaat ctatttaggt ttaaaaattg
     9721 atacagtttt ataagttact ttttcaaaga ctcgtgctgt ctattgcata atgcactgga
     9781 aggagaaaaa aaaggtgcac acgcgtggct ttttcttgaa tttgcagttt gaaaaataac
     9841 tacatggatg ataagaaaac atggagtaca gtcactttga gaaccttcaa tcagctggta
     9901 acgtcttcgt taattggata ctcaaaaaag atggatagca tgaatcacaa gatggaagga
     9961 aatgcgggcc acgaccacag tgatatgcat atgggagatg gagatgatac ctccattggg
    10021 ccgatgaagt tagtcgacgg atagaagcgg ttgtcccctt tcccggcgag ccggcagtcg
    10081 ggccgaggtt cggataaatt ttgtattgtg ttttgattct gtcatgagta ttacttatgt
    10141 tctctttagg taaccccagg ttaatcaatc acagtttcat accggctagt attcaaatta
    10201 tgacttttct tctgcagtgt cagccttacg acgattatct atgagctttg aatatagttt
    10261 gccgtgattc gtatctttaa ttggataata aaatgcgaag gatcgatgac ccttattatt
    10321 atttttctac actggctacc gatttaactc atcttcttga aagtatataa gtaacagtaa
    10381 aatataccgt acttctgcta atgttatttg tcccttattt ttcttttctt gtcttatgct
    10441 atagtaccta agaataacga ctattgtttt gaactaaaca aagtagtaaa agcacataaa
    10501 agaattaaga aaatggctac gctggatgag tcagggatgc ctaagttgtc agagtcgaca
    10561 aaagacggct atctgtgcct taccgtttgt gggtaccgta aaccaggcat gaacgaggaa
    10621 gattaccgca attacatggt caatgtctta gcgcctatgg ccagggacct tatggtgaag
    10681 tacgatatca aacgatggac ccagatccac aatcaaagcg ccactcggga tttaatgaac
    10741 cagctatctg atcatcagat gaacaatatt gcagacttcg actgtttcag ccaggtggtt
    10801 ttcaagagta ttgaagactt caagcgtatg aaacaagacc catggtacaa ggagcattta
    10861 ggcaaccacg gaagttttgc tgatacgaag cgaagcaagg tgacaatagg ctggattgaa
    10921 gaatttgtga gagatggtga ggtagtcgat ggcttcaaga tttgctaggg agattgataa
    10981 gacttttcta gttgcatatc ttttatattt aaatcttatc tattagttaa ttttttgtaa
    11041 tttatcctta tatatagtct ggttattcta aaatatcatt tcagtatcta aaaattcccc
    11101 tcttttttca gttatatctt aacaggcgac agtccaaatg ttgatttatc ccagtccgat
    11161 tcatcagggt tgtgaagcat tttgtcaatg gtcgaaatca catcagtaat agtgcctctt
    11221 acttgcctca tagaatttct ttctcttaac gtcaccgttt ggtcttttat agtttcgaaa
    11281 tctatgtaca acgcaaagaa gctggaaaat tggcgaggga atcctgtttc tgtctatcca
    11341 agaatgggca tgaggtggca accgtcgtgc tagcgtacag ggtgcacttt gtaaccattt
    11401 gggacaccgg acactcgctg ttttcgaaat taccctttaa gcgcgggtat tgaaccaggc
    11461 ttatgcccaa gatcgtagca agcatttaaa tgacggtatc gataagcttg atatcgaatt
    11521 cctgcagccc gggggatcca ctagttctag attaattaa
//
