LOCUS       sequence-307667-        7142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a fungal biosynthetic gene cluster.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7142)
  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 7142)
  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: pCHSU68.2-2.2
FEATURES             Location/Qualifiers
     source          1..7142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      760..947
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(815..839)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3038..3058)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3038..3056)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(3395..3983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3475..3494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4154..4945,4946..5014))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4777..4796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5015..5119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5146..6488
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6109..6156)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6529..6547
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6889..6905)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6889..6905)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aaataagaga atttcagatt gagagaatga aaaaaaaaaa aaaaaaaaag gcagaggaga
       61 gcatagaaat ggggttcact ttttggcaaa gttacagtat gcttattaca tataaataga
      121 gtgccgataa tggctttttt tcatcttcga aatacgcttg ctactgctct tccagcgttt
      181 ttattacttc tttcttgttt ctccttagta tataaaatat caagctacaa caagcataca
      241 atcaactgtc aactgtcaat tatattataa tacactatgt cagagcaacc tacgcccgac
      301 gaggtcttta gctgcctagc tgccttgttt gagtgggctg aaagttatga taccaaggat
      361 tgggaacgtt tgcggcaatg tctcgctccc attctccgcg tggattatcg ccaggtgatg
      421 ggcaagattt gggaagaaat gcttgcagat gaatttattc cgctagcctc gagccctcat
      481 ttcctaggcg atgaactact caggacgcag cactttatcg gcggcgcatc tagctggaac
      541 aaggtatctg ataaggagat acagggccat catcaagtcc gagtcgctca tcaacgctat
      601 acggatagct cgatgaaaga ggtcgcgatt caaggccatg cccacggcgg cgcaacgatg
      661 tggtacaaga aggtcgaagg caaatggaag tttgctgggt tatgtccaaa catcagatgg
      721 ggcgaattta attatgatga gatattcgct ccacggtgag cgaatttctt atgatttatg
      781 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
      841 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
      901 tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat gcctctcaaa
      961 tcttttagcg ccaaggactc caactaattg tatcttgaat ttgcctttac gatccgtttg
     1021 tccagtcacg gcatgtatat cttattaatc ctgcctttct aatcacgtat tctaatgttc
     1081 aattaaggga ttttatcttc atcaacggct cccacgcaaa aaatgacgtt ttgcacacag
     1141 acacgaaata caccttccac cggaacaacg gccatctcca acttataagt tggggaaata
     1201 agacaatttc agacttcaga gaatgaaaaa aaaaaaaggt acatcacaga tggggttcag
     1261 gtttgctaca attgcaggga gcctgtcaca tataaataga cctccagtga tgatatcttt
     1321 cagtcttcaa acgtctcttg tcacagttct ggtcgttcta tatcacatct ctcttggttc
     1381 tacttattgt ctataatatc aagctacagc aagcatacaa tcaactatct accataccat
     1441 aatacacaat ggcattctac ccccccgaca gtctacattt cccgccatta tcacggccgt
     1501 tcctcaactc ttatctgcct ccattattat cagatgacaa ggacacaaca aaggaagatg
     1561 aactatcccg accatttaca accctcacat tcgccacatc tctcgactct tctctatctc
     1621 tggcaccagg tgtgcgcact accctgtctg gacctcaatc aaaagcaatg acacactacc
     1681 tccgcttccg tcacgaagca atcctcatcg gcgtcgggac cgcaatcgca gataatcccg
     1741 gcctgaactg ccgcattgaa ggcgcaagcg gctacgaaga acacccaagc cgggatcccg
     1801 ctattcaggc tctggccgat gaggcgtcat caaaccgcga attgaaagat ctcatgaaag
     1861 ttgttgccaa ctcgaaggcg acgcaagagc aattaaaaac atttcagagc catattgatc
     1921 gggttcaagt agacacgata cgaaaacggc aaaactcact gcatcagccg aggcctataa
     1981 tcattgatcc aaatgctaga tgggtgccca gtgaatctag caaaatcatt cagttagtta
     2041 aacaatataa ggggaaggca ccattcgtcg taacatcaaa atcaacatct ccgcctcagg
     2101 aagctcggca agtcttggag aaatacggtg gaaaatacat ttccttggac actattacca
     2161 gtgataaccc tgatgatgct actggtacac atcaacactt tgactggagt ctcattctca
     2221 acactctagc aacaaaagaa aacatcaaaa gcataatgat cgaaggcggc ggctcaatca
     2281 tcaattcgtt actttcagaa cagcgctata gcactctgat tgactctgtt atcgtgacta
     2341 ttgcgcctac atggttgggc cggggtggcg tggtcgtatc tcccgaaaga cgtgttgacg
     2401 agaatggatg tgcgatacca gcttcgaggc ttacggatgt taaatggtat ccgtttggag
     2461 aagatgtggt tttgtgtggg aggatcaaag gtgttgactg aggagattga taagactttt
     2521 ctagttgcat atcttttata tttaaatctt atctattagt taattttttg taatttatcc
     2581 ttatatatag tctggttatt ctaaaatatc atttcagtat ctaaaaattc ccctcttttt
     2641 tcagttatat cttaacaggc gacagtccaa atgttgattt atcccagtcc gattcatcag
     2701 ggttgtgaag cattttgtca atggtcgaaa tcacatcagt aatagtgcct cttacttgcc
     2761 tcatagaatt tctttctctt aacgtcaccg tttggtcttt tatagtttcg aaatctatgg
     2821 tgataccaaa tggtgttccc aattcatcgt tacgggcgta ttttttacca attgaagtat
     2881 tggaatcgtc aattttaaag tatatctctc ttttacgtaa agcctgcgag atcctcttaa
     2941 gtatagcggg gaagccatcg ttattcgata ttgtcgtaac aaatactttg atcggcgcta
     3001 tgcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     3061 gcttggcgag actcaaacaa gatatatttt gcccgcctta cagacgaaac tagttggagg
     3121 ttatggagca tactatcacg tgggcggcca ctggtgagtt actacacccc aggggcaacg
     3181 ttgatgctcc taaaaaactc tggctggacg caagccgtaa cacccgtgtc acttcataat
     3241 cgtttgcaat tcagggcttg atctacacat taccctgtta tccctattaa ttaatacggt
     3301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga agcatctgtg
     5161 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     5221 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     5281 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     5341 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     5401 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     5461 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     5521 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     5581 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     5641 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     5701 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     5761 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     5821 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     5881 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     5941 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     6001 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     6061 tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga agttcctata
     6121 ctttctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct
     6181 tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct
     6241 gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa
     6301 atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat
     6361 attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct
     6421 atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat
     6481 attggatcat actaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6541 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg agcgcgtttc ggtgatgacg
     6601 gtgaaaacct cggcgcgcca ttaccctgtt atccctaata tacaacgcaa agaagctgga
     6661 aaattggcga gggaatcctg tttctgtcta tccaagaatg ggcatgaggt ggcaaccgtc
     6721 gtgctagcgt acagggtgca ctttgtaacc atttgggaca ccggacactc gctgttttcg
     6781 aaattaccct ttaagcgcgg gtattgaacc aggcttatgc ccaagatcgt agcaagcatt
     6841 taaatgacgg tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtt
     6901 ctagattaat taatatctaa aaattgcctt atgatccgtc tctccggtta cagcctgtgt
     6961 aactgattaa tcctgccttt ctaatcacca ttctaatgtt ttaattaagg gattttgtct
     7021 tcattaacgg ctttcgctca taaaaatgtt atgacgtttt gcccgcaggc gggaaaccat
     7081 ccacttcacg agactgatct cctctgccgg aacaccgggc atctccaact tataagttgg
     7141 ag
//
