LOCUS       sequence-484547-        9529 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Orthogonal PYR1/HAB1 sensor module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9529)
  AUTHORS   Park SY, Qiu J, Wei S, Peterson FC, Beltran J, Medina-Cucurella AV, 
            Vaidya AS, Xing Z, Volkman BF, Nusinow DA, Whitehead TA, Wheeldon I,
            Cutler SR
  TITLE     An orthogonalized PYR1-based CID module with reprogrammable 
            ligand-binding specificity.
  JOURNAL   Nat Chem Biol. 2024 Jan;20(1):103-110. doi: 
            10.1038/s41589-023-01447-7. Epub 2023 Oct 23.
  PUBMED    37872402
REFERENCE   2  (bases 1 to 9529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-023-01447-7"; journalName: "Nat Chem Biol"; date: 
            "2024-01"; volume: "20"; issue: "1"; pages: "103-110"
COMMENT     Sequence Label: pACT-HAB1_star
FEATURES             Location/Qualifiers
     source          1..9529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      668..855
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(723..747)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(876..909)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1418..2512)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(2513..2917)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      3394..4736
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4357..4404)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5094..5112)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5180..5284
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5285..5353,5354..6145)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5503..5522)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6316..6904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6805..6824
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7058..7075
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7090..7230
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7156..7175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7316..7338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        7367..7768
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             7814..7834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7850..8191
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
ORIGIN
        1 aacaaccaag aagtctgcga aatagcaagg agacggatat tgatgtggca caagaagaac
       61 ggtgcaccgc ctctagcaga gagaggcaaa ggaatagatc cagcttgcca agccgcagct
      121 gactacctct caatgcttgc tctacaaaaa ggaagtaaag acaacatctc catcattgtg
      181 attgacttga aagctcaagc aaagttcaag accagaacct gaagcttaat tacaagcagt
      241 acgcttactt actttttttt ttactggggg gagaggtaaa tctcgagaga tctatgaatc
      301 gtagatactg aaaaaccccg caagttcact tcaactgtgc atcgtgcacc atctcaattt
      361 ctttcattta tacatcgttt tgccttcttt tatgtaacta tactcctcta agtttcaatc
      421 ttggccatgt aacctctgat ctatagaatt ttttaaatga ctagaattaa tgcccatctt
      481 ttttttggac ctaaattctt catgaaaata tattacgagg gcttattcag aagctttgga
      541 cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt ctaccttgcc
      601 agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg ttgacacttc
      661 taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
      721 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
      781 taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga
      841 ccacacctct accggcatgc cggtcgaggg acctaataac ttcgtatagc atacattata
      901 cgaagttata ttaagggtta ttgaatatga tcggaattgg tcgactacgt cgttaaggcc
      961 gtttctgaca gagtaaaatt cttgagggaa ctttcaccat tatgggaaat ggttcaagaa
     1021 ggtattgact taaactccat caaatggtca ggtcattgag tgttttttat ttgttgtatt
     1081 tttttttttt tagagaaaat cctccaatat ataaattagg aatcatagtt tcatgatttt
     1141 ctgttacacc taactttttg tgtggtgccc tcctccttgt caatattaat gttaaagtgc
     1201 aattcttttt ccttatcacg ttgagccatt agtatcaatt tgcttacctg tattccttta
     1261 catcctcctt tttctccttc ttgataaatg tatgtagatt gcgtatatag tttcgtctac
     1321 cctatgaaca tattccattt tgtaatttcg tgtcgtttct attatgaatt tcatttataa
     1381 agtttatgta caaatatcat aaaaaaagag aatcttttta agcaaggatt ttcttaactt
     1441 cttcggcgac agcatcaccg acttcggtgg tactgttgga accacctaaa tcaccagttc
     1501 tgatacctgc atccaaaacc tttttaactg catcttcaat ggccttacct tcttcaggca
     1561 agttcaatga caatttcaac atcattgcag cagacaagat agtggcgata gggttgacct
     1621 tattctttgg caaatctgga gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt
     1681 tcttgtctgg caaagaggcc aaggacgcag atggcaacaa acccaaggaa cctgggataa
     1741 cggaggcttc atcggagatg atatcaccaa acatgttgct ggtgattata ataccattta
     1801 ggtgggttgg gttcttaact aggatcatgg cggcagaatc aatcaattga tgttgaacct
     1861 tcaatgtagg gaattcgttc ttgatggttt cctccacagt ttttctccat aatcttgaag
     1921 aggccaaaac attagcttta tccaaggacc aaataggcaa tggtggctca tgttgtaggg
     1981 ccatgaaagc ggccattctt gtgattcttt gcacttctgg aacggtgtat tgttcactat
     2041 cccaagcgac accatcacca tcgtcttcct ttctcttacc aaagtaaata cctcccacta
     2101 attctctgac aacaacgaag tcagtacctt tagcaaattg tggcttgatt ggagataagt
     2161 ctaaaagaga gtcggatgca aagttacatg gtcttaagtt ggcgtacaat tgaagttctt
     2221 tacggatttt tagtaaacct tgttcaggtc taacactacc ggtaccccat ttaggaccac
     2281 ccacagcacc taacaaaacg gcatcagcct tcttggaggc ttccagcgcc tcatctggaa
     2341 gtggaacacc tgtagcatcg atagcagcac caccaattaa atgattttcg aaatcgaact
     2401 tgacattgga acgaacatca gaaatagctt taagaacctt aatggcttcg gctgtgattt
     2461 cttgaccaac gtggtcacct ggcaaaacga cgatcttctt aggggcagac attagaatgg
     2521 tatatccttg aaatatatat atatattgct gaaatgtaaa aggtaagaaa agttagaaag
     2581 taagacgatt gctaaccacc tattggaaaa aacaataggt ccttaaataa tattgtcaac
     2641 ttcaagtatt gtgatgcaag catttagtca tgaacgcttc tctattctat atgaaaagcc
     2701 ggttccggcg ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc
     2761 gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat
     2821 agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga
     2881 actcttgcat cttacgatac tgagtattcc cacagttcct taccactctt ttgttactct
     2941 attgatccag ctcagcaaag gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa
     3001 ctagctagac cgagaaagag actagaaatg caaaaggcac ttctacaatg gctgccatca
     3061 ttattatccg atgtgacgct gcagcttctc aatgatattc gaatacgctt tgaggagata
     3121 cagcctaata tccgacaaac tgttttacag atttacgatc gtacttgtta cccatcattg
     3181 aattttgaac atccgaacct gggagttttc cctgaaacag atagtatatt tgaacctgta
     3241 taataatata tagtctagcg ctttacggaa gacaatgtat gtatttcggt tcctggagaa
     3301 actattgcat ctattgcata ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt
     3361 ttccatcttg cacttcaata gcatatcttt gttaacgaag catctgtgct tcattttgta
     3421 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3481 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt
     3541 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     3601 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     3661 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     3721 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     3781 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     3841 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     3901 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     3961 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     4021 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     4081 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     4141 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     4201 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     4261 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg
     4321 cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagagaa
     4381 taggaacttc ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca
     4441 acgcgagctg cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg
     4501 tatatatata tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat
     4561 atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc
     4621 catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac
     4681 tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat tggatcatat
     4741 gcatagtacc gagaaactag tgcgaagtag tgatcaggta ttgctgttat ctgatgagta
     4801 tacgttgtcc tggccacggc agaagcacgc ttatcgctcc aatttcccac aacattagtc
     4861 aactccgtta ggcccttcat tgaaagaaat gaggtcatca aatgtcttcc aatgtgagat
     4921 tttgggccat tttttatagc aaagattgaa taaggcgcat ttttcttcaa agctttattg
     4981 tacgatctga ctaagttatc ttttaataat tggtattcct gtttattgct tgaagaattg
     5041 ccggtcctat ttactcgttt taggactggt tcagaattct tgaagacgaa agggcctcgt
     5101 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5161 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5221 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5281 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5341 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5401 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5461 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5521 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5581 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5641 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5701 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5761 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5821 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5881 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5941 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6001 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6061 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6121 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6181 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6241 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6421 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6961 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7021 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7081 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7141 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     7201 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     7261 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7321 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcaacttctt
     7381 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
     7441 aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt
     7501 tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattca
     7561 cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga
     7621 aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt
     7681 ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt
     7741 tcaagctata ccaagcatac aatcaactcc aagctttgca aagatggata aagcggaatt
     7801 aattcccgag cctccaaaaa agaagagaaa ggtcgaattg ggtaccgccg ccaattttaa
     7861 tcaaagtggg aatattgctg atagctcatt gtccttcact ttcactaaca gtagcaacgg
     7921 tccgaacctc ataacaactc aaacaaattc tcaagcgctt tcacaaccaa ttgcctcctc
     7981 taacgttcat gataacttca tgaataatga aatcacggct agtaaaattg atgatggtaa
     8041 taattcaaaa ccactgtcac ctggttggac ggaccaaact gcgtataacg cgtttggaat
     8101 cactacaggg atgtttaata ccactacaat ggatgatgta tataactatc tattcgatga
     8161 tgaagatacc ccaccaaacc caaaaaaaga gatctggaat tcggatcctc gagatttacc
     8221 tgagatgact cccgcagttg caatgactct tagcttagca gccaacacca tgtgtgaatc
     8281 atcacctgtc gagatcactc agctaaagaa cgttactgat gcagctgact tgttatctga
     8341 ttctgaaaat caaagctttt gcaacggagg gactgaatgc actatggaag atgtttctga
     8401 actggaagag gtaggtgaac aggatttgtt gaaaacttta tccgatacga gaagcgggtc
     8461 ttccaatgtt tttgatgaag acgatgtatt gtctgttgtg gaggataata gtgctgtcat
     8521 aagtgagggc ttgttagttg ttgatgcagg ctctgaatta agcttgtcta atacagctat
     8581 ggaaatagat aacgggcgag ttcttgcaac cgcgattatc gtaggcgaat caagcattga
     8641 gcaggttccc accgcggaag ttcttatcgc gggtgtaaat caggatacca atacttcgga
     8701 ggttgtcatt agattgccag atgaaaatag taatcatctg gtgaaaggga gaagtgttta
     8761 tgaactagat tgtataccgc tttggggcac ggtttccatt caagggaatg cgtctgagat
     8821 ggaggctgct tttgccgtgt cacctcattt tctgaaacta cccatcaaaa tgcttatggg
     8881 ggaccatgag ggtatgagtc caagcctcac acacctcaca ggtcattttt tcggtgttta
     8941 tgatggtcat ggaggccata aggttgctga ctattgccga gatagactcc attttgcttt
     9001 ggctgaagaa atagaacgta taaaagacga attatgcaag aggaatacag gagagggtag
     9061 gcaggtgcag tgggataaag tcttcacgag ttgttttcta actgtcgatg gtgagattga
     9121 aggaaaaatt ggtagagccg ttgttggttc ttctgataag gttcttgagg ctgttgcgga
     9181 tgagaccgta ggatcaactg ctgttgttgc cttggtttgc tcatcacata tagtagtttc
     9241 taactgcggt gattcgaggg cggttttatt ccgtggcaaa gaagccatgc ccttgtcagt
     9301 tgatcacaaa ccagatagag aggatgaata tgcaagaata gaaaatgctg gaggcaaagt
     9361 tatacaatgg caaggcgcac gtgtttttgg tcgtctcgcc atgtctaggt ccatcggtga
     9421 cagatatctg aagccatatg tgatcccaga accggaagtg acattcatgc ctcggtcaag
     9481 agaagacgag tgtctcatac tagccagtga cggtctttgg gatgtaatg
//
