LOCUS       sequence-363425-       10021 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Biosensor for acetic acid: BM3R1-Haa1-mTurquoise2 under HAA1p and 
            mCherry under BM3R1ENO1cp with flanks for genome integration in toHO
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10021)
  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 10021)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_9
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(54..869)
                     /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(54..869)
                     /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     777..796
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     861..880
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1094..1682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1583..1602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7456..8166
                     /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(7601..7619)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7869..7888
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8630..9433
                     /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 gagttagttc tgccccgagc ggccgcgtgt tacaaccaat taaccaattc tgattagaaa
       61 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
      121 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
      181 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
      241 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
      301 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
      361 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
      421 gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
      481 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
      541 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
      601 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
      661 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
      721 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
      781 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag
      841 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
      901 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
      961 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagtca
     1021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1201 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1741 tgttctttcc tgcgttatcc cctgattctg tggataaccg tgcggccgcc aattctaatt
     1801 gtatcgagat cacttttcgt gatccgctaa tcagcgacgg tcacattagg tttgccaagt
     1861 cagggtatga accatacgat cagttttcgt gaacctggta cgtatattgt ggcgtttgtg
     1921 tatattttca ttctttgaca acaatcaata ccaacctcaa ataggaaaag taataagttt
     1981 ggcgttacac cccaaaagac gccaaacgga tcgaacttac tcaatagcaa ttagcgagac
     2041 aaaacctacg ttaagacctg taaccgattt atcaaagcac tctgcggttc tttcttggga
     2101 atattacctg gacattttgt gccctcaaga aacgaggctc tacgagcctg ttggagcccc
     2161 tcagacatta gccgccacga atcaaacttt ttacgcgatt cggcccaaat cagtttctca
     2221 cagatcattc gtagagtgaa aaagcacatc gattatttga tacccctttg ggttaattac
     2281 tgttgaggtc tttgccgtaa gttccatcgc tgtccctgaa ttcgcatcta gactgaactg
     2341 gccgataatt gcagacgaac gccttgcaat tagcttctat tacacctgat gactgttttt
     2401 caatctttat gtgatctttt tacacccaat gtgataaagt actagtaaca tgaatactac
     2461 taaacagatg atactttaga gttttcattc caacactcaa ttgtaaaaat aagacaatat
     2521 tatgattttt tggctgtata accacatcta tttagtagca gggtgacatg ctactcccac
     2581 acaatacttc gcgcaagtgg tttagtggta aaatccaacg ttgccatcgt tgggcccccg
     2641 gttcgattcc gggcttgcgc aatttttccc cctcgccctg aagggcgtaa cgttaacgct
     2701 tccggtaatt tgcgaataat ttttcttccc atttcccctt tcttttccct ttaacatgtg
     2761 gagcatagca ggctctttac ttttttacgc attcagacag tcgggaggtc aattggtggt
     2821 gccggttgtg ggaaagaggt aattttattt cctttattgg acatgatttc aatagtcaca
     2881 ttggggcagg aaaaagaaaa gggagaaaaa aattcagtaa agaaaaaaaa taagaaaaag
     2941 agaaaagaaa caagagaggg ctacagattt ctaaatatcc ttttgctgca cattttttat
     3001 ttacctgcta attggtctta gtaatacaga gtcgtgcatt tccactttat aagaaaaaaa
     3061 aaagggataa cagcaccagc acttgattga aaactagaaa gttcacgttt tataagtcct
     3121 ttccgttctc tttatcagtt ataataaaag agccaagaca aaacgtggca gggaagcttt
     3181 tccaaggaaa agagggttgt aatattaaac aaaaaccgta atcaagcaaa ataagacccc
     3241 gtggtttttt atcataggaa aagagttctc tgttgtcact ctaccatatt tgacaagaaa
     3301 aaggaaacaa aagtatagaa aaaaaaaacc taaaaaatat atggaatcta ctcctactaa
     3361 gcaaaaagcc atcttctctg cctccttgtt gttgttcgct gagagaggtt tcgacgctac
     3421 cactatgcca atgattgccg aaaacgctaa ggttggtgct ggtactatct acagatactt
     3481 caagaacaaa gaatccttag ttaatgagtt gtttcaacaa cacgtcaatg aatttttgca
     3541 atgtatcgaa tctggtttgg ctaacgaaag agatggttac agggacggtt tccaccacat
     3601 cttcgaaggt atggtcacct tcaccaagaa ccatcctaga gccttaggtt tcatcaagac
     3661 ccactctcaa ggtacttttt tgaccgaaga atccagattg gcttatcaaa agttggtcga
     3721 atttgtctgt actttcttca gagaaggtca aaagcaaggt gtcatcagaa atttgccaga
     3781 aaacgctttg attgctatct tgttcggttc tttcatggaa gtctacgaaa tgattgaaaa
     3841 cgattatttg tctttgactg atgaattatt aaccggtgtt gaggaatcct tgtgggctgc
     3901 cttgtctcgt caatctggtt ctatggtctt gataaatggc ataaagtatg cctgtgagag
     3961 gtgcataaga ggccatagag taacaacatg caatcataca gatcaaccgc ttatgatgat
     4021 caaacccaaa ggtagacctt ccactacatg cgactattgt aaacaacttc gaaaaaacaa
     4081 gaatgcaaat cctgaaggtg tttgcacgtg tggccggcta gagaagaaaa aactggcaca
     4141 gaaagccaaa gaagaagcaa gagctaaagc caaagaaaaa caaagaaaac agtgtacctg
     4201 cgggactgat gaggtttgca aatatcatgc tcaaaagaga catctaagaa agtccccttc
     4261 aagttctcaa aagaaaggaa gatccatttc tcgttctcaa ccaatgtttg aaagggtatt
     4321 gtcttctact tcacttgaca gcaatatgtt atccggccac ggagcactat cagatacctc
     4381 tagcatactg acgagcacat ttttagacag tgagccgggt gttggtaaaa tttcaaaaga
     4441 ttaccatcat gtcccttcat tggcctccat ttcatcctta caatcctcgc aatcgttaga
     4501 tcaaaatttc agtataccac aaagcccgcc gttatcttca atgtcattta attttctcac
     4561 gggaaatatc aatgaaacca accaaaatca cagtaatcat cagcattcaa aatcaggcaa
     4621 taactggcaa gatagttcgg taagcttgcc agcgaaagct gattcacgtc ttaacatgat
     4681 ggataaaaac aactctgtgg gtcttgacct attaggccat tcaaaacgaa tatcgccgat
     4741 atcaaactct cgtgtgggcg aagttagcgt tccgctagaa gaatatattc cttctgacat
     4801 tgatggggtt ggaagagtta ctgataaaag ctctttggtc tacgattggc catttgatga
     4861 aagtattgag agaaatttca gtacaaccgc aaccgctgca actggtgaaa gtaagttcga
     4921 cattaacgac aactgtaata gaattaatag caaaagttat agtaagacta atagtatgaa
     4981 tggaaacggt atgaacaata gcaataataa taatatcaac agtaatggca acgacaagaa
     5041 caataacaac tcttctagac aagaacatca aggaaatgga ctatttgaca tgtttacaga
     5101 ttcatcgtcg atttcaacgc tttcccgtgc aaacttatta ttgcaagaaa aaattggttc
     5161 gcaagaaaac tctgtcaaac aagaaaacta ttcgaaaaat cctcaacttc gtcatcaatt
     5221 aacttccaga agtagatcat ttattcatca tccggcaaac gagtatttga agaatacttt
     5281 tggaaattca catagtaatg acatcggaaa gggagttgaa gtgctatctt tgacaccgag
     5341 ttttatggat attcccgaaa aagaaagaga aacggaaaga tcgccatcat ccaattacat
     5401 tactgacaga cctttcactc gaaaacctag atcttctagc attgacgtaa accataggta
     5461 tccacctatg gcaccaacaa ccgtagcgac atctcccggt gcattgaaca atgccgtagc
     5521 aagcaatctc gacgatcaac tgagtttaac atcactaaac tctcagccat catcgatagc
     5581 aaatatgatg atggaccctt caaacctagc tgagcaaagt tctattcatt cagttcctca
     5641 gtcaataaac tctccgagaa tgcctaaaac tggaagtcgc caagacaaga acattcacac
     5701 taagaaggaa gaaagaaatc cgctaaataa catacacgat ctgtcacaat tggaaaatgt
     5761 accagacgag atgaaccaaa tgttctcccc accattaaaa agtatgaata gaccggatgc
     5821 cataagggaa aattcatcta gtagtaattt cataatccaa ggaaatagca tgatctctac
     5881 gccttccgga aggaatgacc ttccagatac ctctccaatg agtagtattc aaacagcgtc
     5941 accaccaagt caattactga ccgatcaagg atttgcggat ttggataatt tcatgtcttc
     6001 gttaggttct gtttctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     6061 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     6121 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
     6181 gccaacctta gtcactactt tatcttgggg tgttcaatgt tttgcaagat acccagatca
     6241 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     6301 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     6361 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     6421 aggtcacaaa ttggaataca attatttctc tgacaatgtt tacatcactg ctgacaaaca
     6481 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca
     6541 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     6601 caaccattac ttatccactc aatctaagtt atccaaagat ccaaacgaaa agagggacca
     6661 catggtcttg ttagaatttg ttactgctgc tggtattacc ttgggtatgg atgaattgta
     6721 caaaggatcc taactcgaga gcttttgatt aagccttcta gtccaaaaaa cacgtttttt
     6781 tgtcatttat ttcattttct tagaatagtt tagtttattc attttatagt cacgaatgtt
     6841 ttatgattct atatagggtt gcaaacaagc atttttcatt ttatgttaaa acaatttcag
     6901 gtttaccttt tattctgctt gtggtgacgc gtgtatccgc ccgctctttt ggtcacccat
     6961 gtatgctgcc aaaccagatg tcaacacagc tacaacggct agtcggaatg aacattcatt
     7021 ccgtgaccta ggatgtgacg gaatgaaggt tcattccgga ctctagataa gcacggaatg
     7081 aactttcatt ccgctgaagc ttgtcaatcg gaatgaaggt tcattccggc tagtcggaat
     7141 gaacattcat tccgcgacct aggatgtgac ggaatgaagg ttcattccgg actctagata
     7201 agcacggaat gaactttcat tccgctgaag cttgtcaatc ggaatgaagg ttcattccgg
     7261 ctatctcccc ggaaactgtg gccttttctg gcacacatga tctccacgat ttcaacatat
     7321 aaatagcttt tgataatggc aatattaatc aaatttattt tacttctttc ttgtaacatc
     7381 tctcttgtaa tcccttattc cttctagcta tttttcataa aaaaccaagc aactgcttat
     7441 caacacacaa acactatggt gagcaagggc gaggaggata acatggccat catcaaggag
     7501 ttcatgcgct tcaaggtgca catggagggc tccgtggacg gccacgagtt cgagatcgag
     7561 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     7621 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     7681 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     7741 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     7801 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     7861 tccgacggcc ccgtaatgca gaagaagact atgggctggg aggcctcctc cgagcggatg
     7921 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     7981 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     8041 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     8101 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     8161 aagtaaatcc taactcgagg ccaattggtg cggcaattga taataacgaa aatgtctttt
     8221 aatgatctgg gtataatgag gaattttccg aacgttttta ctttatatat atatatacat
     8281 gtaacatata ttctatacgc tatagagaaa ggaaattttt caattaaaaa aaaaatagag
     8341 aaagagtttc acttcttgat tatcgctaac actaatggtt gaagtactgc tactttaatt
     8401 ttatgctgag cagttacaga gatgttacga accactagtg cactgcagta cacggtttcc
     8461 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     8521 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     8581 cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc
     8641 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     8701 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     8761 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     8821 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     8881 caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     8941 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     9001 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     9061 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     9121 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     9181 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     9241 gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt
     9301 ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     9361 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     9421 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     9481 agcttcaatt taattatatc agttattacc cgagttatcg tgttgcatct gcggctttaa
     9541 attgatgtat ctcatcgcag gcacgggcag tacagtgccc tgagcgtagg gaaaaatgaa
     9601 aaaaaggatg taacttttaa cataattcca gcacgcagcg attgggtata atgaagattg
     9661 ttaagttcaa caacatgtga aattcttagc tccgcaactt gcttgaatta tgagctctaa
     9721 gattcaagaa gtaaataggt catataatgg aatgatagta gtacttgata aaggaaccat
     9781 gtgatcttac gttgatatga aaataattct ctcacagaag tcagtttgta tgactaacat
     9841 agagagtaat tgcttttcgg ataaaatcgc acctaacaaa gattaccttt tgtaaagcct
     9901 ccagaacagc tatgaatgtt cgttattcta cgcactttct ttcatacatc ttgatccgcc
     9961 ttttttgatg taggttttaa cgcggcaaaa catattaatt tttctgattt ggaaaaataa
    10021 a
//
