LOCUS       sequence-235539-        4040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  low copy plasmid, neg. control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4040)
  AUTHORS   Steiger MG, Rassinger A, Mattanovich D, Sauer M
  TITLE     Engineering of the citrate exporter protein enables high citric acid
            production in Aspergillus niger.
  JOURNAL   Metab Eng. 2019 Mar;52:224-231. doi: 10.1016/j.ymben.2018.12.004. 
            Epub 2018 Dec 13.
  PUBMED    30553933
REFERENCE   2  (bases 1 to 4040)
  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: "Metab 
            Eng."; date: "2019-03"; volume: "52:224-231. doi"; pages: " 
            10.1016/j.ymben.2018.12.004. Epub 2018 Dec 13"
COMMENT     Sequence Label: BB3_arscen_URA3_Hygro
FEATURES             Location/Qualifiers
     source          1..4040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1..1576
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1..344
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             345..1373
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             345..1373
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1379..1576
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1588..2091
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      complement(2154..2742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2234..2253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2840..3055
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3056..3859
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
       61 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      121 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacgtgcgag
      181 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      241 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      301 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaaaagcctg
      361 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
      421 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
      481 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
      541 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
      601 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctccctg
      661 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
      721 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
      781 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
      841 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
      901 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
      961 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     1021 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     1081 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     1141 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     1201 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     1261 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     1321 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatcagtac
     1381 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     1441 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     1501 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     1561 gctggtcgct atactggctt ccttttcatc acgtgctata aaaataatta taatttaaat
     1621 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     1681 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     1741 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     1801 acaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     1861 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     1921 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     1981 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     2041 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat cgcgctgtga
     2101 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2161 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2221 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2281 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2341 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2401 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2461 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2521 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2581 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2641 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2701 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2761 tctacggggt ggagcgcaaa aaaccccgcc cctgacaggg cggggttttt tcgcgatcgg
     2821 agcagggtcc ataaagcttt tcaattcatc tttttttttt tattcttttt tttgattccg
     2881 gtttccttga aatttttttg attcggtaat ctccgagcag aaggaagaac gaaggaagga
     2941 gcacagactt agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc
     3001 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
     3061 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
     3121 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
     3181 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
     3241 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
     3301 caagtacaat tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa
     3361 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
     3421 cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcggaag aagtaacaaa
     3481 ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc tagctactgg
     3541 agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt
     3601 tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc
     3661 cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa ccgtggatga
     3721 tgtggtgtct acaggatctg acattattat tgttggaaga ggactatttg caaagggaag
     3781 ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag
     3841 atgcggccag caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca
     3901 acttagagct tcaatttaat tatatcagtt attacccggg aatctcggcg ctccggggag
     3961 gcgtcttcgg atccctcgag gaagacgccg ctaattggag gcgtcttcgg atccctcgag
     4021 gaagacgccg ctagctcatg
//
