LOCUS       sequence-363416-        9358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for biosensor in pMM2_9: HAA1-mTurquoise2 under HAA1p and 
            mCherry under HREsENO1cp with flanks for genome integration into HO 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9358)
  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 9358)
  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_11
FEATURES             Location/Qualifiers
     source          1..9358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(61..876)
                     /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(61..876)
                     /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     784..803
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     868..887
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1101..1689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1590..1609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6800..7510
                     /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(6945..6963)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7213..7232
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7974..8777
                     /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 aaataaagag ttagttctgc cccgagcggc cgcgtgttac aaccaattaa ccaattctga
       61 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
      121 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      181 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      241 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      301 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
      361 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
      421 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
      481 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
      541 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
      601 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
      661 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      721 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
      781 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
      841 cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
      901 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
      961 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     1021 tcagtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc ggccgccaat
     1801 tctaattgta tcgagatcac ttttcgtgat ccgctaatca gcgacggtca cattaggttt
     1861 gccaagtcag ggtatgaacc atacgatcag ttttcgtgaa cctggtacgt atattgtggc
     1921 gtttgtgtat attttcattc tttgacaaca atcaatacca acctcaaata ggaaaagtaa
     1981 taagtttggc gttacacccc aaaagacgcc aaacggatcg aacttactca atagcaatta
     2041 gcgagacaaa acctacgtta agacctgtaa ccgatttatc aaagcactct gcggttcttt
     2101 cttgggaata ttacctggac attttgtgcc ctcaagaaac gaggctctac gagcctgttg
     2161 gagcccctca gacattagcc gccacgaatc aaacttttta cgcgattcgg cccaaatcag
     2221 tttctcacag atcattcgta gagtgaaaaa gcacatcgat tatttgatac ccctttgggt
     2281 taattactgt tgaggtcttt gccgtaagtt ccatcgctgt ccctgaattc gcatctagac
     2341 tgaactggcc gataattgca gacgaacgcc ttgcaattag cttctattac acctgatgac
     2401 tgtttttcaa tctttatgtg atctttttac acccaatgtg ataaagtact agtaacatga
     2461 atactactaa acagatgata ctttagagtt ttcattccaa cactcaattg taaaaataag
     2521 acaatattat gattttttgg ctgtataacc acatctattt agtagcaggg tgacatgcta
     2581 ctcccacaca atacttcgcg caagtggttt agtggtaaaa tccaacgttg ccatcgttgg
     2641 gcccccggtt cgattccggg cttgcgcaat ttttccccct cgccctgaag ggcgtaacgt
     2701 taacgcttcc ggtaatttgc gaataatttt tcttcccatt tcccctttct tttcccttta
     2761 acatgtggag catagcaggc tctttacttt tttacgcatt cagacagtcg ggaggtcaat
     2821 tggtggtgcc ggttgtggga aagaggtaat tttatttcct ttattggaca tgatttcaat
     2881 agtcacattg gggcaggaaa aagaaaaggg agaaaaaaat tcagtaaaga aaaaaaataa
     2941 gaaaaagaga aaagaaacaa gagagggcta cagatttcta aatatccttt tgctgcacat
     3001 tttttattta cctgctaatt ggtcttagta atacagagtc gtgcatttcc actttataag
     3061 aaaaaaaaaa gggataacag caccagcact tgattgaaaa ctagaaagtt cacgttttat
     3121 aagtcctttc cgttctcttt atcagttata ataaaagagc caagacaaaa cgtggcaggg
     3181 aagcttttcc aaggaaaaga gggttgtaat attaaacaaa aaccgtaatc aagcaaaata
     3241 agaccccgtg gttttttatc ataggaaaag agttctctgt tgtcactcta ccatatttga
     3301 caagaaaaag gaaacaaaag tatagaaaaa aaaaacctaa aaaatatatg atggtcttga
     3361 taaatggcat aaagtatgcc tgtgagaggt gcataagagg ccatagagta acaacatgca
     3421 atcatacaga tcaaccgctt atgatgatca aacccaaagg tagaccttcc actacatgcg
     3481 actattgtaa acaacttcga aaaaacaaga atgcaaatcc tgaaggtgtt tgcacgtgtg
     3541 gccggctaga gaagaaaaaa ctggcacaga aagccaaaga agaagcaaga gctaaagcca
     3601 aagaaaaaca aagaaaacag tgtacctgcg ggactgatga ggtttgcaaa tatcatgctc
     3661 aaaagagaca tctaagaaag tccccttcaa gttctcaaaa gaaaggaaga tccatttctc
     3721 gttctcaacc aatgtttgaa agggtattgt cttctacttc acttgacagc aatatgttat
     3781 ccggccacgg agcactatca gatacctcta gcatactgac gagcacattt ttagacagtg
     3841 agccgggtgt tggtaaaatt tcaaaagatt accatcatgt cccttcattg gcctccattt
     3901 catccttaca atcctcgcaa tcgttagatc aaaatttcag tataccacaa agcccgccgt
     3961 tatcttcaat gtcatttaat tttctcacgg gaaatatcaa tgaaaccaac caaaatcaca
     4021 gtaatcatca gcattcaaaa tcaggcaata actggcaaga tagttcggta agcttgccag
     4081 cgaaagctga ttcacgtctt aacatgatgg ataaaaacaa ctctgtgggt cttgacctat
     4141 taggccattc aaaacgaata tcgccgatat caaactctcg tgtgggcgaa gttagcgttc
     4201 cgctagaaga atatattcct tctgacattg atggggttgg aagagttact gataaaagct
     4261 ctttggtcta cgattggcca tttgatgaaa gtattgagag aaatttcagt acaaccgcaa
     4321 ccgctgcaac tggtgaaagt aagttcgaca ttaacgacaa ctgtaataga attaatagca
     4381 aaagttatag taagactaat agtatgaatg gaaacggtat gaacaatagc aataataata
     4441 atatcaacag taatggcaac gacaagaaca ataacaactc ttctagacaa gaacatcaag
     4501 gaaatggact atttgacatg tttacagatt catcgtcgat ttcaacgctt tcccgtgcaa
     4561 acttattatt gcaagaaaaa attggttcgc aagaaaactc tgtcaaacaa gaaaactatt
     4621 cgaaaaatcc tcaacttcgt catcaattaa cttccagaag tagatcattt attcatcatc
     4681 cggcaaacga gtatttgaag aatacttttg gaaattcaca tagtaatgac atcggaaagg
     4741 gagttgaagt gctatctttg acaccgagtt ttatggatat tcccgaaaaa gaaagagaaa
     4801 cggaaagatc gccatcatcc aattacatta ctgacagacc tttcactcga aaacctagat
     4861 cttctagcat tgacgtaaac cataggtatc cacctatggc accaacaacc gtagcgacat
     4921 ctcccggtgc attgaacaat gccgtagcaa gcaatctcga cgatcaactg agtttaacat
     4981 cactaaactc tcagccatca tcgatagcaa atatgatgat ggacccttca aacctagctg
     5041 agcaaagttc tattcattca gttcctcagt caataaactc tccgagaatg cctaaaactg
     5101 gaagtcgcca agacaagaac attcacacta agaaggaaga aagaaatccg ctaaataaca
     5161 tacacgatct gtcacaattg gaaaatgtac cagacgagat gaaccaaatg ttctccccac
     5221 cattaaaaag tatgaataga ccggatgcca taagggaaaa ttcatctagt agtaatttca
     5281 taatccaagg aaatagcatg atctctacgc cttccggaag gaatgacctt ccagatacct
     5341 ctccaatgag tagtattcaa acagcgtcac caccaagtca attactgacc gatcaaggat
     5401 ttgcggattt ggataatttc atgtcttcgt taggttctgt ttctaaaggt gaagaattat
     5461 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
     5521 ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt
     5581 gtactactgg taaattgcca gttccatggc caaccttagt cactacttta tcttggggtg
     5641 ttcaatgttt tgcaagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
     5701 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
     5761 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
     5821 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaat tatttctctg
     5881 acaatgttta catcactgct gacaaacaaa agaatggtat caaagctaac ttcaaaatta
     5941 gacacaacat tgaagatggt ggtgttcaat tagctgacca ttatcaacaa aatactccaa
     6001 ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa tctaagttat
     6061 ccaaagatcc aaacgaaaag agggaccaca tggtcttgtt agaatttgtt actgctgctg
     6121 gtattacctt gggtatggat gaattgtaca aaggatccta actcgagagc ttttgattaa
     6181 gccttctagt ccaaaaaaca cgtttttttg tcatttattt cattttctta gaatagttta
     6241 gtttattcat tttatagtca cgaatgtttt atgattctat atagggttgc aaacaagcat
     6301 ttttcatttt atgttaaaac aatttcaggt ttacctttta ttctgcttgt ggtgacgcgt
     6361 gtatccgccc gctcttttgg tcacccatgt atgctgccaa accagatgtc aacacagcta
     6421 caacggctag tggcgagggg tgacctagga tgtgaggcga gggggactct agataagcag
     6481 gcgaggggct gaagcttgtc aatggcgagg gggctagtgg cgaggggcga cctaggatgt
     6541 gaggcgaggg ggactctaga taagcaggcg aggggctgaa gcttgtcaat ggcgaggggg
     6601 ctatctcccc ggaaactgtg gccttttctg gcacacatga tctccacgat ttcaacatat
     6661 aaatagcttt tgataatggc aatattaatc aaatttattt tacttctttc ttgtaacatc
     6721 tctcttgtaa tcccttattc cttctagcta tttttcataa aaaaccaagc aactgcttat
     6781 caacacacaa acacgctata tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     6841 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg gacggccacg agttcgagat
     6901 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     6961 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     7021 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     7081 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     7141 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     7201 cccctccgac ggccccgtaa tgcagaagaa gactatgggc tgggaggcct cctccgagcg
     7261 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     7321 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     7381 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     7441 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     7501 gtacaagtaa atcctaactc gaggccaatt ggtgcggcaa ttgataataa cgaaaatgtc
     7561 ttttaatgat ctgggtataa tgaggaattt tccgaacgtt tttactttat atatatatat
     7621 acatgtaaca tatattctat acgctataga gaaaggaaat ttttcaatta aaaaaaaaat
     7681 agagaaagag tttcacttct tgattatcgc taacactaat ggttgaagta ctgctacttt
     7741 aattttatgc tgagcagtta cagagatgtt acgaaccact agtgcactgc agtacacggt
     7801 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     7861 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     7921 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     7981 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     8041 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     8101 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     8161 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     8221 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     8281 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     8341 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     8401 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     8461 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     8521 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     8581 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     8641 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     8701 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     8761 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     8821 ttagagcttc aatttaatta tatcagttat tacccgagtt atcgtgttgc atctgcggct
     8881 ttaaattgat gtatctcatc gcaggcacgg gcagtacagt gccctgagcg tagggaaaaa
     8941 tgaaaaaaag gatgtaactt ttaacataat tccagcacgc agcgattggg tataatgaag
     9001 attgttaagt tcaacaacat gtgaaattct tagctccgca acttgcttga attatgagct
     9061 ctaagattca agaagtaaat aggtcatata atggaatgat agtagtactt gataaaggaa
     9121 ccatgtgatc ttacgttgat atgaaaataa ttctctcaca gaagtcagtt tgtatgacta
     9181 acatagagag taattgcttt tcggataaaa tcgcacctaa caaagattac cttttgtaaa
     9241 gcctccagaa cagctatgaa tgttcgttat tctacgcact ttctttcata catcttgatc
     9301 cgcctttttt gatgtaggtt ttaacgcggc aaaacatatt aatttttctg atttggaa
//
