LOCUS       sequence-363439-        9866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for biosensor in pMM2_9: HAA1-mTurquoise2 under HAA1p and 
            mCherry under HREsYGP1p with flanks for genome integration into HO 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9866)
  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 9866)
  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_10
FEATURES             Location/Qualifiers
     source          1..9866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(33..848)
                     /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(33..848)
                     /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     756..775
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     840..859
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1073..1661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1562..1581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7280..7990
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVDGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7425..7443)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7693..7712
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8454..9257
                     /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 gccgcgtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga
       61 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
      121 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
      181 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
      241 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta
      301 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
      361 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa tacgcgatcg
      421 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
      481 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
      541 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
      601 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
      661 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
      721 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
      781 aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc ccgttgaata
      841 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
      901 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt
      961 tgaataaatc gaacttttgc tgagttgaag gatcagtcat gaccaaaatc ccttaacgtg
     1021 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1201 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1681 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1741 ctgattctgt ggataaccgt gcggccgcca attctaattg tatcgagatc acttttcgtg
     1801 atccgctaat cagcgacggt cacattaggt ttgccaagtc agggtatgaa ccatacgatc
     1861 agttttcgtg aacctggtac gtatattgtg gcgtttgtgt atattttcat tctttgacaa
     1921 caatcaatac caacctcaaa taggaaaagt aataagtttg gcgttacacc ccaaaagacg
     1981 ccaaacggat cgaacttact caatagcaat tagcgagaca aaacctacgt taagacctgt
     2041 aaccgattta tcaaagcact ctgcggttct ttcttgggaa tattacctgg acattttgtg
     2101 ccctcaagaa acgaggctct acgagcctgt tggagcccct cagacattag ccgccacgaa
     2161 tcaaactttt tacgcgattc ggcccaaatc agtttctcac agatcattcg tagagtgaaa
     2221 aagcacatcg attatttgat acccctttgg gttaattact gttgaggtct ttgccgtaag
     2281 ttccatcgct gtccctgaat tcgcatctag actgaactgg ccgataattg cagacgaacg
     2341 ccttgcaatt agcttctatt acacctgatg actgtttttc aatctttatg tgatcttttt
     2401 acacccaatg tgataaagta ctagtaacat gaatactact aaacagatga tactttagag
     2461 ttttcattcc aacactcaat tgtaaaaata agacaatatt atgatttttt ggctgtataa
     2521 ccacatctat ttagtagcag ggtgacatgc tactcccaca caatacttcg cgcaagtggt
     2581 ttagtggtaa aatccaacgt tgccatcgtt gggcccccgg ttcgattccg ggcttgcgca
     2641 atttttcccc ctcgccctga agggcgtaac gttaacgctt ccggtaattt gcgaataatt
     2701 tttcttccca tttccccttt cttttccctt taacatgtgg agcatagcag gctctttact
     2761 tttttacgca ttcagacagt cgggaggtca attggtggtg ccggttgtgg gaaagaggta
     2821 attttatttc ctttattgga catgatttca atagtcacat tggggcagga aaaagaaaag
     2881 ggagaaaaaa attcagtaaa gaaaaaaaat aagaaaaaga gaaaagaaac aagagagggc
     2941 tacagatttc taaatatcct tttgctgcac attttttatt tacctgctaa ttggtcttag
     3001 taatacagag tcgtgcattt ccactttata agaaaaaaaa aagggataac agcaccagca
     3061 cttgattgaa aactagaaag ttcacgtttt ataagtcctt tccgttctct ttatcagtta
     3121 taataaaaga gccaagacaa aacgtggcag ggaagctttt ccaaggaaaa gagggttgta
     3181 atattaaaca aaaaccgtaa tcaagcaaaa taagaccccg tggtttttta tcataggaaa
     3241 agagttctct gttgtcactc taccatattt gacaagaaaa aggaaacaaa agtatagaaa
     3301 aaaaaaacct aaaaaatata tgatggtctt gataaatggc ataaagtatg cctgtgagag
     3361 gtgcataaga ggccatagag taacaacatg caatcataca gatcaaccgc ttatgatgat
     3421 caaacccaaa ggtagacctt ccactacatg cgactattgt aaacaacttc gaaaaaacaa
     3481 gaatgcaaat cctgaaggtg tttgcacgtg tggccggcta gagaagaaaa aactggcaca
     3541 gaaagccaaa gaagaagcaa gagctaaagc caaagaaaaa caaagaaaac agtgtacctg
     3601 cgggactgat gaggtttgca aatatcatgc tcaaaagaga catctaagaa agtccccttc
     3661 aagttctcaa aagaaaggaa gatccatttc tcgttctcaa ccaatgtttg aaagggtatt
     3721 gtcttctact tcacttgaca gcaatatgtt atccggccac ggagcactat cagatacctc
     3781 tagcatactg acgagcacat ttttagacag tgagccgggt gttggtaaaa tttcaaaaga
     3841 ttaccatcat gtcccttcat tggcctccat ttcatcctta caatcctcgc aatcgttaga
     3901 tcaaaatttc agtataccac aaagcccgcc gttatcttca atgtcattta attttctcac
     3961 gggaaatatc aatgaaacca accaaaatca cagtaatcat cagcattcaa aatcaggcaa
     4021 taactggcaa gatagttcgg taagcttgcc agcgaaagct gattcacgtc ttaacatgat
     4081 ggataaaaac aactctgtgg gtcttgacct attaggccat tcaaaacgaa tatcgccgat
     4141 atcaaactct cgtgtgggcg aagttagcgt tccgctagaa gaatatattc cttctgacat
     4201 tgatggggtt ggaagagtta ctgataaaag ctctttggtc tacgattggc catttgatga
     4261 aagtattgag agaaatttca gtacaaccgc aaccgctgca actggtgaaa gtaagttcga
     4321 cattaacgac aactgtaata gaattaatag caaaagttat agtaagacta atagtatgaa
     4381 tggaaacggt atgaacaata gcaataataa taatatcaac agtaatggca acgacaagaa
     4441 caataacaac tcttctagac aagaacatca aggaaatgga ctatttgaca tgtttacaga
     4501 ttcatcgtcg atttcaacgc tttcccgtgc aaacttatta ttgcaagaaa aaattggttc
     4561 gcaagaaaac tctgtcaaac aagaaaacta ttcgaaaaat cctcaacttc gtcatcaatt
     4621 aacttccaga agtagatcat ttattcatca tccggcaaac gagtatttga agaatacttt
     4681 tggaaattca catagtaatg acatcggaaa gggagttgaa gtgctatctt tgacaccgag
     4741 ttttatggat attcccgaaa aagaaagaga aacggaaaga tcgccatcat ccaattacat
     4801 tactgacaga cctttcactc gaaaacctag atcttctagc attgacgtaa accataggta
     4861 tccacctatg gcaccaacaa ccgtagcgac atctcccggt gcattgaaca atgccgtagc
     4921 aagcaatctc gacgatcaac tgagtttaac atcactaaac tctcagccat catcgatagc
     4981 aaatatgatg atggaccctt caaacctagc tgagcaaagt tctattcatt cagttcctca
     5041 gtcaataaac tctccgagaa tgcctaaaac tggaagtcgc caagacaaga acattcacac
     5101 taagaaggaa gaaagaaatc cgctaaataa catacacgat ctgtcacaat tggaaaatgt
     5161 accagacgag atgaaccaaa tgttctcccc accattaaaa agtatgaata gaccggatgc
     5221 cataagggaa aattcatcta gtagtaattt cataatccaa ggaaatagca tgatctctac
     5281 gccttccgga aggaatgacc ttccagatac ctctccaatg agtagtattc aaacagcgtc
     5341 accaccaagt caattactga ccgatcaagg atttgcggat ttggataatt tcatgtcttc
     5401 gttaggttct gtttctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     5461 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     5521 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
     5581 gccaacctta gtcactactt tatcttgggg tgttcaatgt tttgcaagat acccagatca
     5641 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     5701 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     5761 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     5821 aggtcacaaa ttggaataca attatttctc tgacaatgtt tacatcactg ctgacaaaca
     5881 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca
     5941 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     6001 caaccattac ttatccactc aatctaagtt atccaaagat ccaaacgaaa agagggacca
     6061 catggtcttg ttagaatttg ttactgctgc tggtattacc ttgggtatgg atgaattgta
     6121 caaaggatcc taactcgaga gcttttgatt aagccttcta gtccaaaaaa cacgtttttt
     6181 tgtcatttat ttcattttct tagaatagtt tagtttattc attttatagt cacgaatgtt
     6241 ttatgattct atatagggtt gcaaacaagc atttttcatt ttatgttaaa acaatttcag
     6301 gtttaccttt tattctgctt gtggtgacgc gtgtatccgc ccgctctttt ggtcacccat
     6361 gtatgctgcc aaaccagatg tcaacacagc tacaacggag caataggaaa attcagtagt
     6421 cgatttccat ggcgaggggc tattgggtaa tgtggcggcg agggctaaac attcggtatc
     6481 attgattcaa tgttctcaca atgttcaaat tctcggatcg ggtatcccta attgggatct
     6541 ctttccagag gcgaggggct tgatgtggga ttgaagacac atgagttttt ttctcaccgt
     6601 cgaaactttt gcttacccgt aaaacatcgg ccattcgttg gtttattatg tgactttttt
     6661 gtgacttttt tttttgcgaa gttttctcgt catccgcggg gcgaggggcg ggcgaaaaag
     6721 tctctcactt ttacccaatc ggggcatgtt atgtggacat gttatgtgtc caagagacac
     6781 atgagcccaa aaagggtaca ctgtctaaat agtttaccct ttacgaatcg aaacttctcg
     6841 ggcgaggggc cgaaaattag cgcagaagtt gacgaaagtt tatacttttc ctaatggaga
     6901 ggctcaggag ctgggcgagg ggatattcgt actctattgc atcttcaaac tccgaagaat
     6961 ctcaggcgag ggggaagccg gcttctcgat gctacagttc aacatcgata ctttgaagaa
     7021 agaaagcgcc tattatatct cttttaccct ggcgagggga attaggaggc gaggggaaaa
     7081 aagtacctca ctaaaaaaac catcatctct gaaatataaa aagcttgata gagggtgacg
     7141 gcgaggggaa cttctgctgt gttctcttgg gttattgctc ttattgaata tggcgagggg
     7201 ttctttcttg cttgtaaaaa atcagctcaa aaaacatcta caggattaat cgtcagttaa
     7261 gtaatacagt aatagaaata tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     7321 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg gacggccacg agttcgagat
     7381 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     7441 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     7501 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     7561 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     7621 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     7681 cccctccgac ggccccgtaa tgcagaagaa gactatgggc tgggaggcct cctccgagcg
     7741 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     7801 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     7861 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     7921 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     7981 gtacaagtaa atcctaactc gaggccaatt ggtgcggcaa ttgataataa cgaaaatgtc
     8041 ttttaatgat ctgggtataa tgaggaattt tccgaacgtt tttactttat atatatatat
     8101 acatgtaaca tatattctat acgctataga gaaaggaaat ttttcaatta aaaaaaaaat
     8161 agagaaagag tttcacttct tgattatcgc taacactaat ggttgaagta ctgctacttt
     8221 aattttatgc tgagcagtta cagagatgtt acgaaccact agtgcactgc agtacacggt
     8281 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     8341 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     8401 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     8461 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     8521 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     8581 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     8641 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     8701 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     8761 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     8821 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     8881 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     8941 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     9001 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     9061 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     9121 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     9181 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     9241 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     9301 ttagagcttc aatttaatta tatcagttat tacccgagtt atcgtgttgc atctgcggct
     9361 ttaaattgat gtatctcatc gcaggcacgg gcagtacagt gccctgagcg tagggaaaaa
     9421 tgaaaaaaag gatgtaactt ttaacataat tccagcacgc agcgattggg tataatgaag
     9481 attgttaagt tcaacaacat gtgaaattct tagctccgca acttgcttga attatgagct
     9541 ctaagattca agaagtaaat aggtcatata atggaatgat agtagtactt gataaaggaa
     9601 ccatgtgatc ttacgttgat atgaaaataa ttctctcaca gaagtcagtt tgtatgacta
     9661 acatagagag taattgcttt tcggataaaa tcgcacctaa caaagattac cttttgtaaa
     9721 gcctccagaa cagctatgaa tgttcgttat tctacgcact ttctttcata catcttgatc
     9781 cgcctttttt gatgtaggtt ttaacgcggc aaaacatatt aatttttctg atttggaaaa
     9841 ataaagagtt agttctgccc cgagcg
//
