LOCUS       sequence-235993-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTEF1-cexA-tCYC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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-1313
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1572..1819
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1588..1606)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        1856..2072
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2073..2876
                     /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"
     gene            3018..4593
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        3018..3361
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3362..4390
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             3362..4390
                     /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      4396..4593
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      5190..6067
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6130..6718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6210..6229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6799..7205
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 agtgactccg ataagggatc cctctctccg gagccttcga ccctcgaggc tcagtcatcc
       61 gagaagccac cgcatcatat cttcacacga tctcgcaagc tgcaaatggt ttgcatcgtc
      121 tccctcgctg ccatattttc tccgctttcg tcgaacattt acttccctgc cctggatgat
      181 gtctcgaaat ccctcaacat cagcatgtcg ctcgcaacac tcaccatcac ggtgtacatg
      241 atcgtccaag gcctcgctcc cagcttctgg ggttccatgt cagacgccac aggtagacgg
      301 cctgtcttta ttggaacatt cattgtttac ctcgtagcca atattgctct ggccgaatcc
      361 aagaactatg gtgagctcat ggccttccga gccttgcagg ctgctggtag cgcagccacc
      421 atctcaatcg gagctggagt gattggtgat atcacaaact cggaagaaag aggtagcttg
      481 gtgggtatct tcggtggagt tcgcatgctt ggacagggaa tcgggccggt tttcggcggc
      541 attttcaccc agtatctcgg atatcgatct atcttttggt tcctcacgat tgctggaggc
      601 gtgagtctcc tgtccattct ggtgcttctt ccggagacat tgaggccaat tgctggaaat
      661 ggaactgtga agctcaatgg cattcataag cccttcatct acacgatcac cggccagacg
      721 ggggttgtcg agggagcgca accggaagcg aaaaagacca aaaccagctg gaagtctgtt
      781 tttgctcctt tgacattcct cgtcgaaaag gacgttttca tcaccctgtt ctttggaagt
      841 atcgtgtaca cagtgtggag catggtgaca tccagtacca ccgacctctt cagcgaagtg
      901 tacggcctgt catccctgga cattggactc actttcctag gcaatggctt tggatgtatg
      961 tctggctctt atctggtcgg ctaccttatg gattacaacc accgtcttac cgaacgcgaa
     1021 tattgcgaga aacacggtta tccggcaggc acacgtgtca atctgaaatc acaccccgac
     1081 ttccccattg aggtcgcccg gatgcgcaat acctggtggg tgattgcgat cttcatcgtg
     1141 acagttgctt tgtacggcgt gtctttgcgg acacatctgg cggtgcctat cattctgcag
     1201 tacttcattg cgttctgctc aacaggactc ttcaccatca acagcgccct ggtcatcgat
     1261 ctttacccag gtgctagcgc cagtgcgaca gcagtgaaca atctgatgcg gtgcctgctt
     1321 ggagctggcg gtgtggctat cgtgcaacct atcctggacg ccttgaagcc ggattatact
     1381 ttcctcttgc ttgccggcat cacccttgtg atgactccgt tgctgtacgt cgaagatcga
     1441 tggggtcctg gctggcgaca tgcccgcgaa aggagactca aggccaaagc caacggcaac
     1501 taggcttcgt ccgacggcgg cccgacgggt ccgaggcctc ggagatccgt cccccttttc
     1561 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     1621 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1681 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     1741 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     1801 ctcgaaggct ttaatttgca agctggcgct ccggggagca gggtccataa agcttttcaa
     1861 ttcatctttt ttttttttgt tctttttttt gattccggtt tctttgaaat ttttttgatt
     1921 cggtaatctc cgagcagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1981 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     2041 gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     2101 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     2161 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     2221 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     2281 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     2341 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     2401 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     2461 ttagcggttt gaagcaggcg gcggaagaag taacaaagga acctagaggc cttttgatgt
     2521 tagcagaatt gtcatgcaag ggctccctag ctactggaga atatactaag ggtactgttg
     2581 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     2641 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     2701 gagacgcatt gggtcaacag tatagaaccg tggatgatgt ggtgtctaca ggatctgaca
     2761 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     2821 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2881 ctgtattata agtaaatgca tgtatactaa actcacaact tagagcttca atttaattat
     2941 atcagttatt acccgggaat ctcggcgcta attggaggcg tcttcggatc cctcgaggaa
     3001 gacgccgcta gctcatggac atggaggccc agaataccct ccttgacagt cttgacgtgc
     3061 gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat
     3121 aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct
     3181 cgctgcagac gtgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg
     3241 ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata
     3301 tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac
     3361 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3421 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     3481 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     3541 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     3601 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     3661 gttgcaagac ctccctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     3721 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     3781 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     3841 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     3901 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     3961 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4021 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4081 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4141 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4201 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4261 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     4321 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     4381 aaaggaataa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     4441 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     4501 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     4561 gtcaatcgta tgtgaatgct ggtcgctata ctggctttta tcgatgataa gctgtcaaac
     4621 atgagaatta attccacgga ctatagacta tacctagtat actccgtcta ctgtacgata
     4681 cacttccgct caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg
     4741 atccagctca gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag
     4801 ctagaccgag aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat
     4861 tatccgatgt gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc
     4921 ctaatatccg acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt
     4981 ttgaacatcc gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat
     5041 aatatatagt ctagcgcttt acggaagaga atgtatgtat ttcggttcct ggagaaacta
     5101 ttgcatctat tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc
     5161 atcttgcact tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac
     5221 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     5281 aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt
     5341 aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     5401 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     5461 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     5521 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     5581 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     5641 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     5701 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     5761 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     5821 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     5881 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     5941 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     6001 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     6061 tcgcaatgcg ctgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6721 agatcctttg atcttttcta cggggtggag cgcaaaaaac cccgcccctg acagggcggg
     6781 gttttttcgc gatcggagca cacaccatag cttcaaaatg tttctactcc ttttttactc
     6841 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca
     6901 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg
     6961 gaaaagaaaa aagcgaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt
     7021 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct
     7081 cccattgata tttaagttaa taaacggtgt tcaatttctc aagtttcagt ttcatttttc
     7141 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat
     7201 ctaagggcgg tcatgtcctc aaccacgtcc tcatcaagat cagaccttga aaaggtcccc
     7261 gtaccacagg tcacccctag agac
//
