LOCUS       sequence-363440-        9868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HAA1-mTurquoise2 under HAA1p and mRuby2 under YGP1p with flanks for 
            genome integration into HO locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9868)
  AUTHORS   Mormino M, Siewers V, Nygard Y
  TITLE     Development of an Haa1-based biosensor for acetic acid sensing in 
            Saccharomyces cerevisiae.
  JOURNAL   FEMS Yeast Res. 2021 Sep 11;21(6). pii: 6363685. doi: 
            10.1093/femsyr/foab049.
  PUBMED    34477863
REFERENCE   2  (bases 1 to 9868)
  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: "FEMS Yeast 
            Res."; date: "2021-09-11"; volume: "21(6). pii: 6363685. doi"; 
            pages: " 10.1093/femsyr/foab049"
COMMENT     Sequence Label: pMM2_4
FEATURES             Location/Qualifiers
     source          1..9868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(16..831)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(16..831)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     739..758
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     823..842
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1056..1644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1545..1564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7263..7973
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             8439..9242
                     /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"
ORIGIN
        1 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
       61 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
      121 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
      181 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
      241 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
      301 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
      361 attcattcgt gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt
      421 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
      481 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
      541 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
      601 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
      661 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
      721 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
      781 ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct
      841 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
      901 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
      961 tgctgagttg aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga
     1021 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1081 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1141 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1201 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1261 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1321 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1381 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1441 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1501 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1561 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1621 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1681 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1741 cgtgcggccg ccaattctaa ttgtatcgag atcacttttc gtgatccgct aatcagcgac
     1801 ggtcacatta ggtttgccaa gtcagggtat gaaccatacg atcagttttc gtgaacctgg
     1861 tacgtatatt gtggcgtttg tgtatatttt cattctttga caacaatcaa taccaacctc
     1921 aaataggaaa agtaataagt ttggcgttac accccaaaag acgccaaacg gatcgaactt
     1981 actcaatagc aattagcgag acaaaaccta cgttaagacc tgtaaccgat ttatcaaagc
     2041 actctgcggt tctttcttgg gaatattacc tggacatttt gtgccctcaa gaaacgaggc
     2101 tctacgagcc tgttggagcc cctcagacat tagccgccac gaatcaaact ttttacgcga
     2161 ttcggcccaa atcagtttct cacagatcat tcgtagagtg aaaaagcaca tcgattattt
     2221 gatacccctt tgggttaatt actgttgagg tctttgccgt aagttccatc gctgtccctg
     2281 aattcgcatc tagactgaac tggccgataa ttgcagacga acgccttgca attagcttct
     2341 attacacctg atgactgttt ttcaatcttt atgtgatctt tttacaccca atgtgataaa
     2401 gtactagtaa catgaatact actaaacaga tgatacttta gagttttcat tccaacactc
     2461 aattgtaaaa ataagacaat attatgattt tttggctgta taaccacatc tatttagtag
     2521 cagggtgaca tgctactccc acacaatact tcgcgcaagt ggtttagtgg taaaatccaa
     2581 cgttgccatc gttgggcccc cggttcgatt ccgggcttgc gcaatttttc cccctcgccc
     2641 tgaagggcgt aacgttaacg cttccggtaa tttgcgaata atttttcttc ccatttcccc
     2701 tttcttttcc ctttaacatg tggagcatag caggctcttt acttttttac gcattcagac
     2761 agtcgggagg tcaattggtg gtgccggttg tgggaaagag gtaattttat ttcctttatt
     2821 ggacatgatt tcaatagtca cattggggca ggaaaaagaa aagggagaaa aaaattcagt
     2881 aaagaaaaaa aataagaaaa agagaaaaga aacaagagag ggctacagat ttctaaatat
     2941 ccttttgctg cacatttttt atttacctgc taattggtct tagtaataca gagtcgtgca
     3001 tttccacttt ataagaaaaa aaaaagggat aacagcacca gcacttgatt gaaaactaga
     3061 aagttcacgt tttataagtc ctttccgttc tctttatcag ttataataaa agagccaaga
     3121 caaaacgtgg cagggaagct tttccaagga aaagagggtt gtaatattaa acaaaaaccg
     3181 taatcaagca aaataagacc ccgtggtttt ttatcatagg aaaagagttc tctgttgtca
     3241 ctctaccata tttgacaaga aaaaggaaac aaaagtatag aaaaaaaaaa cctaaaaaat
     3301 atatgatggt cttgataaat ggcataaagt atgcctgtga gaggtgcata agaggccata
     3361 gagtaacaac atgcaatcat acagatcaac cgcttatgat gatcaaaccc aaaggtagac
     3421 cttccactac atgcgactat tgtaaacaac ttcgaaaaaa caagaatgca aatcctgaag
     3481 gtgtttgcac gtgtggccgg ctagagaaga aaaaactggc acagaaagcc aaagaagaag
     3541 caagagctaa agccaaagaa aaacaaagaa aacagtgtac ctgcgggact gatgaggttt
     3601 gcaaatatca tgctcaaaag agacatctaa gaaagtcccc ttcaagttct caaaagaaag
     3661 gaagatccat ttctcgttct caaccaatgt ttgaaagggt attgtcttct acttcacttg
     3721 acagcaatat gttatccggc cacggagcac tatcagatac ctctagcata ctgacgagca
     3781 catttttaga cagtgagccg ggtgttggta aaatttcaaa agattaccat catgtccctt
     3841 cattggcctc catttcatcc ttacaatcct cgcaatcgtt agatcaaaat ttcagtatac
     3901 cacaaagccc gccgttatct tcaatgtcat ttaattttct cacgggaaat atcaatgaaa
     3961 ccaaccaaaa tcacagtaat catcagcatt caaaatcagg caataactgg caagatagtt
     4021 cggtaagctt gccagcgaaa gctgattcac gtcttaacat gatggataaa aacaactctg
     4081 tgggtcttga cctattaggc cattcaaaac gaatatcgcc gatatcaaac tctcgtgtgg
     4141 gcgaagttag cgttccgcta gaagaatata ttccttctga cattgatggg gttggaagag
     4201 ttactgataa aagctctttg gtctacgatt ggccatttga tgaaagtatt gagagaaatt
     4261 tcagtacaac cgcaaccgct gcaactggtg aaagtaagtt cgacattaac gacaactgta
     4321 atagaattaa tagcaaaagt tatagtaaga ctaatagtat gaatggaaac ggtatgaaca
     4381 atagcaataa taataatatc aacagtaatg gcaacgacaa gaacaataac aactcttcta
     4441 gacaagaaca tcaaggaaat ggactatttg acatgtttac agattcatcg tcgatttcaa
     4501 cgctttcccg tgcaaactta ttattgcaag aaaaaattgg ttcgcaagaa aactctgtca
     4561 aacaagaaaa ctattcgaaa aatcctcaac ttcgtcatca attaacttcc agaagtagat
     4621 catttattca tcatccggca aacgagtatt tgaagaatac ttttggaaat tcacatagta
     4681 atgacatcgg aaagggagtt gaagtgctat ctttgacacc gagttttatg gatattcccg
     4741 aaaaagaaag agaaacggaa agatcgccat catccaatta cattactgac agacctttca
     4801 ctcgaaaacc tagatcttct agcattgacg taaaccatag gtatccacct atggcaccaa
     4861 caaccgtagc gacatctccc ggtgcattga acaatgccgt agcaagcaat ctcgacgatc
     4921 aactgagttt aacatcacta aactctcagc catcatcgat agcaaatatg atgatggacc
     4981 cttcaaacct agctgagcaa agttctattc attcagttcc tcagtcaata aactctccga
     5041 gaatgcctaa aactggaagt cgccaagaca agaacattca cactaagaag gaagaaagaa
     5101 atccgctaaa taacatacac gatctgtcac aattggaaaa tgtaccagac gagatgaacc
     5161 aaatgttctc cccaccatta aaaagtatga atagaccgga tgccataagg gaaaattcat
     5221 ctagtagtaa tttcataatc caaggaaata gcatgatctc tacgccttcc ggaaggaatg
     5281 accttccaga tacctctcca atgagtagta ttcaaacagc gtcaccacca agtcaattac
     5341 tgaccgatca aggatttgcg gatttggata atttcatgtc ttcgttaggt tctgtttcta
     5401 aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta
     5461 atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga
     5521 ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta
     5581 ctttatcttg gggtgttcaa tgttttgcaa gatacccaga tcatatgaaa caacatgact
     5641 ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg
     5701 acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa
     5761 tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat
     5821 acaattattt ctctgacaat gtttacatca ctgctgacaa acaaaagaat ggtatcaaag
     5881 ctaacttcaa aattagacac aacattgaag atggtggtgt tcaattagct gaccattatc
     5941 aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca
     6001 ctcaatctaa gttatccaaa gatccaaacg aaaagaggga ccacatggtc ttgttagaat
     6061 ttgttactgc tgctggtatt accttgggta tggatgaatt gtacaaagga tcctaactcg
     6121 agagcttttg attaagcctt ctagtccaaa aaacacgttt ttttgtcatt tatttcattt
     6181 tcttagaata gtttagttta ttcattttat agtcacgaat gttttatgat tctatatagg
     6241 gttgcaaaca agcatttttc attttatgtt aaaacaattt caggtttacc ttttattctg
     6301 cttgtggtga cgcgtgtatc cgcccgctct tttggtcacc catgtatgct gccaaaccag
     6361 atgtcaacac agctacaacg gagcaatagg aaaattcagt agtcgatttc catttcaatt
     6421 ccctattggg taatgtggcg gcgagggcta aacattcggt atcattgatt caatgttctc
     6481 acaatgttca aattctcgga tcgggtatcc ctaattggga tctctttcca gaggaagggg
     6541 ccttgatgtg ggattgaaga cacatgagtt tttttctcac cgtcgaaact tttgcttacc
     6601 cgtaaaacat cggccattcg ttggtttatt atgtgacttt tttgtgactt ttttttttgc
     6661 gaagttttct cgtcatccgc ggaagatccg gcgggcgaaa aagtctctca cttttaccca
     6721 atcggggcat gttatgtgga catgttatgt gtccaagaga cacatgagcc caaaaagggt
     6781 acactgtcta aatagtttac cctttacgaa tcgaaacttc tcgaagaagg ggccgaaaat
     6841 tagcgcagaa gttgacgaaa gtttatactt ttcctaatgg agaggctcag gagctgaaaa
     6901 gggatatatt cgtactctat tgcatcttca aactccgaag aatctcagta gggagtgaag
     6961 ccggcttctc gatgctacag ttcaacatcg atactttgaa gaaagaaagc gcctattata
     7021 tctcttttac cctattagta ataattagga aaaagggaga aaaaagtacc tcactaaaaa
     7081 aaccatcatc tctgaaatat aaaaagcttg atagagggtg acatttgcta gaacttctgc
     7141 tgtgttctct tgggttattg ctcttattga atatcccttc tatttctttc ttgcttgtaa
     7201 aaaatcagct caaaaaacat ctacaggatt aatcgtcagt taagtaatac agtaatagaa
     7261 atatggtgtc caaaggagag gagttaatca aggaaaacat gagaatgaaa gttgtcatgg
     7321 agggctccgt taatggtcac caattcaagt gtacagggga aggtgaaggt aatccttaca
     7381 tgggtacaca aactatgaga attaaagtaa ttgaaggcgg accactacca tttgcatttg
     7441 acattctggc aacgtcattc atgtacggat cacgaacttt catcaagtac cctaaaggta
     7501 taccagactt tttcaagcaa tcttttccag agggttttac atgggaaagg gttacaagat
     7561 acgaagatgg gggtgtcgtc acagttatgc aagatacttc attagaagat ggctgccttg
     7621 tctatcatgt gcaagtaaga ggggtgaatt ttccttctaa cggacctgtg atgcagaaaa
     7681 agaccaaagg ttgggaacca aatactgaaa tgatgtaccc agctgatgga ggtttgagag
     7741 gctacacaca catggcgctt aaagttgatg gtggaggtca tttgtcttgt agttttgtta
     7801 ccacttatcg ttctaaaaag actgttggca atatcaaaat gccaggaata catgctgtag
     7861 accacagact agaaagactc gaagagagcg ataacgaaat gttcgttgta cagagagagc
     7921 atgccgtagc caaatttgct ggcttaggcg gtggtatgga tgaattgtat aagggatcct
     7981 aactcgaggc caattggtgc ggcaattgat aataacgaaa atgtctttta atgatctggg
     8041 tataatgagg aattttccga acgtttttac tttatatata tatatacatg taacatatat
     8101 tctatacgct atagagaaag gaaatttttc aattaaaaaa aaaatagaga aagagtttca
     8161 cttcttgatt atcgctaaca ctaatggttg aagtactgct actttaattt tatgctgagc
     8221 agttacagag atgttacgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     8281 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     8341 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     8401 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     8461 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     8521 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     8581 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     8641 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     8701 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     8761 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     8821 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     8881 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     8941 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     9001 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     9061 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     9121 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     9181 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     9241 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     9301 aattatatca gttattaccc gagttatcgt gttgcatctg cggctttaaa ttgatgtatc
     9361 tcatcgcagg cacgggcagt acagtgccct gagcgtaggg aaaaatgaaa aaaaggatgt
     9421 aacttttaac ataattccag cacgcagcga ttgggtataa tgaagattgt taagttcaac
     9481 aacatgtgaa attcttagct ccgcaacttg cttgaattat gagctctaag attcaagaag
     9541 taaataggtc atataatgga atgatagtag tacttgataa aggaaccatg tgatcttacg
     9601 ttgatatgaa aataattctc tcacagaagt cagtttgtat gactaacata gagagtaatt
     9661 gcttttcgga taaaatcgca cctaacaaag attacctttt gtaaagcctc cagaacagct
     9721 atgaatgttc gttattctac gcactttctt tcatacatct tgatccgcct tttttgatgt
     9781 aggttttaac gcggcaaaac atattaattt ttctgatttg gaaaaataaa gagttagttc
     9841 tgccccgagc ggccgcgtgt tacaacca
//
