LOCUS       sequence-235546-        7519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTPI1-cexA-tCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7519)
  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 7519)
  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: pMST-1312
FEATURES             Location/Qualifiers
     source          1..7519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            909..2484
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        909..1252
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1253..2281
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1253..2281
                     /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      2287..2484
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      3081..3958
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(4021..4609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4828..5336
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     terminator      6982..7229
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6998..7016)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        7266..7482
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             join(7483..7519,1..767)
                     /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 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
       61 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
      121 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
      181 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
      241 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
      301 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
      361 tgaagcaggc ggcggaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
      421 tgtcatgcaa gggctcccta gctactggag aatatactaa gggtactgtt gacattgcga
      481 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
      541 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
      601 tgggtcaaca gtatagaacc gtggatgatg tggtgtctac aggatctgac attattattg
      661 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
      721 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
      781 aagtaaatgc atgtatacta aactcacaac ttagagcttc aatttaatta tatcagttat
      841 tacccgggaa tctcggcgct aattggaggc gtcttcggat ccctcgagga agacgccgct
      901 agctcatgga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
      961 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     1021 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     1081 cgtgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     1141 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     1201 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
     1261 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     1321 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     1381 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     1441 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     1501 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     1561 cctccctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     1621 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     1681 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     1741 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     1801 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     1861 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     1921 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     1981 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     2041 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     2101 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     2161 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     2221 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     2281 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     2341 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     2401 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     2461 atgtgaatgc tggtcgctat actggctttt atcgatgata agctgtcaaa catgagaatt
     2521 aattccacgg actatagact atacctagta tactccgtct actgtacgat acacttccgc
     2581 tcaggtcctt gtcctttaac gaggccttac cactcttttg ttactctatt gatccagctc
     2641 agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta gctagaccga
     2701 gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta ttatccgatg
     2761 tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag cctaatatcc
     2821 gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat tttgaacatc
     2881 cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa taatatatag
     2941 tctagcgctt tacggaagag aatgtatgta tttcggttcc tggagaaact attgcatcta
     3001 ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac
     3061 ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     3121 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3181 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     3241 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3301 gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta
     3361 caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct
     3421 cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg
     3481 ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt
     3541 cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc
     3601 cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt
     3661 gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact
     3721 acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta
     3781 ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga
     3841 gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac
     3901 agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatgc
     3961 gctgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4441 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4621 gatcttttct acggggtgga gcgcaaaaaa ccccgcccct gacagggcgg ggttttttcg
     4681 cgatcggagg aatccatcaa tagatacgtc ctgaggaccg tgctacccaa atggactgat
     4741 tgtgagggag agctaactac atagtgttta aagattacgg atatttaact tacttagaat
     4801 aatgccattt ttttgagtta taataatcct acgttagtgt gagcgggatt taaactgtga
     4861 ggaccttaat acattcagac acttctgcgg tatcacccta cttattccct tcgagattat
     4921 atctaggaac ccatcaggtt ggtggaagat tacccgttct aagacttttc agcttcctct
     4981 attgatgtta cacctggaca ccccttttct ggcatccagt ttttaatctt cagtggcatg
     5041 tgagattctc cgaaattaat taaagcaatc acacaattct ctcggatacc acctcggttg
     5101 aaactgacag gtggtttgtt acgcatgcta atgcaaagga gcctatatac ctttggctcg
     5161 gctgctgtaa cagggaatat aaagggcagc ataatttagg agtttagtga acttgcaaca
     5221 tttactattt tcccttctta cgtaaatatt tttcttttta attctaaatc aatctttttc
     5281 aattttttgt ttgtattctt ttcttgctta aatctataac tacaaaaaac acatacacat
     5341 gtcctcaacc acgtcctcat caagatcaga ccttgaaaag gtccccgtac cacaggtcac
     5401 ccctagagac agtgactccg ataagggatc cctctctccg gagccttcga ccctcgaggc
     5461 tcagtcatcc gagaagccac cgcatcatat cttcacacga tctcgcaagc tgcaaatggt
     5521 ttgcatcgtc tccctcgctg ccatattttc tccgctttcg tcgaacattt acttccctgc
     5581 cctggatgat gtctcgaaat ccctcaacat cagcatgtcg ctcgcaacac tcaccatcac
     5641 ggtgtacatg atcgtccaag gcctcgctcc cagcttctgg ggttccatgt cagacgccac
     5701 aggtagacgg cctgtcttta ttggaacatt cattgtttac ctcgtagcca atattgctct
     5761 ggccgaatcc aagaactatg gtgagctcat ggccttccga gccttgcagg ctgctggtag
     5821 cgcagccacc atctcaatcg gagctggagt gattggtgat atcacaaact cggaagaaag
     5881 aggtagcttg gtgggtatct tcggtggagt tcgcatgctt ggacagggaa tcgggccggt
     5941 tttcggcggc attttcaccc agtatctcgg atatcgatct atcttttggt tcctcacgat
     6001 tgctggaggc gtgagtctcc tgtccattct ggtgcttctt ccggagacat tgaggccaat
     6061 tgctggaaat ggaactgtga agctcaatgg cattcataag cccttcatct acacgatcac
     6121 cggccagacg ggggttgtcg agggagcgca accggaagcg aaaaagacca aaaccagctg
     6181 gaagtctgtt tttgctcctt tgacattcct cgtcgaaaag gacgttttca tcaccctgtt
     6241 ctttggaagt atcgtgtaca cagtgtggag catggtgaca tccagtacca ccgacctctt
     6301 cagcgaagtg tacggcctgt catccctgga cattggactc actttcctag gcaatggctt
     6361 tggatgtatg tctggctctt atctggtcgg ctaccttatg gattacaacc accgtcttac
     6421 cgaacgcgaa tattgcgaga aacacggtta tccggcaggc acacgtgtca atctgaaatc
     6481 acaccccgac ttccccattg aggtcgcccg gatgcgcaat acctggtggg tgattgcgat
     6541 cttcatcgtg acagttgctt tgtacggcgt gtctttgcgg acacatctgg cggtgcctat
     6601 cattctgcag tacttcattg cgttctgctc aacaggactc ttcaccatca acagcgccct
     6661 ggtcatcgat ctttacccag gtgctagcgc cagtgcgaca gcagtgaaca atctgatgcg
     6721 gtgcctgctt ggagctggcg gtgtggctat cgtgcaacct atcctggacg ccttgaagcc
     6781 ggattatact ttcctcttgc ttgccggcat cacccttgtg atgactccgt tgctgtacgt
     6841 cgaagatcga tggggtcctg gctggcgaca tgcccgcgaa aggagactca aggccaaagc
     6901 caacggcaac taggcttcgt ccgacggcgg cccgacgggt ccgaggcctc ggagatccgt
     6961 cccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc
     7021 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     7081 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     7141 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     7201 ttttgggacg ctcgaaggct ttaatttgca agctggcgct ccggggagca gggtccataa
     7261 agcttttcaa ttcatctttt ttttttttgt tctttttttt gattccggtt tctttgaaat
     7321 ttttttgatt cggtaatctc cgagcagaag gaagaacgaa ggaaggagca cagacttaga
     7381 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     7441 caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     7501 aaggaacgtg ctgctactc
//
