LOCUS       sequence-484463-        8740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for HAB1 mutants library construction from oligopools; PYR1
            sensor library assembly from oligopools; .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8740)
  TITLE     Cutler lab 2025
REFERENCE   2  (bases 1 to 8740)
  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"
COMMENT     Sequence Label: pACT-HAB1-backbone
FEATURES             Location/Qualifiers
     source          1..8740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      691..878
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(746..770)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(899..932)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1441..2535)
                     /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(2536..2940)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      3417..4759
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4380..4427)
                     /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(5117..5135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5203..5307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5308..5376,5377..6168)
                     /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(5526..5545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6339..6927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6828..6847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7081..7098
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7113..7253
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7179..7198)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7339..7361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        7390..7791
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             7837..7857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7873..8214
                     /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 cagaggagac ctccgaggtc tcgaacaacc aagaagtctg cgaaatagca aggagacgga
       61 tattgatgtg gcacaagaag aacggtgcac cgcctctagc agagagaggc aaaggaatag
      121 atccagcttg ccaagccgca gctgactacc tctcaatgct tgctctacaa aaaggaagta
      181 aagacaacat ctccatcatt gtgattgact tgaaagctca aagaaagttc aagaccagaa
      241 cctgaagctt aattacaagc agtacgctta cttacttttt tttttactgg ggggagaggt
      301 aaatctcgag agatctatga atcgtagata ctgaaaaacc ccgcaagttc acttcaactg
      361 tgcatcgtgc accatctcaa tttctttcat ttatacatcg ttttgccttc ttttatgtaa
      421 ctatactcct ctaagtttca atcttggcca tgtaacctct gatctataga attttttaaa
      481 tgactagaat taatgcccat cttttttttg gacctaaatt cttcatgaaa atatattacg
      541 agggcttatt cagaagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag
      601 gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt
      661 ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt
      721 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
      781 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
      841 tagcatgagg tcgctcttat tgaccacacc tctaccggca tgccggtcga gggacctaat
      901 aacttcgtat agcatacatt atacgaagtt atattaaggg ttattgaata tgatcggaat
      961 tggtcgacta cgtcgttaag gccgtttctg acagagtaaa attcttgagg gaactttcac
     1021 cattatggga aatggttcaa gaaggtattg acttaaactc catcaaatgg tcaggtcatt
     1081 gagtgttttt tatttgttgt attttttttt ttttagagaa aatcctccaa tatataaatt
     1141 aggaatcata gtttcatgat tttctgttac acctaacttt ttgtgtggtg ccctcctcct
     1201 tgtcaatatt aatgttaaag tgcaattctt tttccttatc acgttgagcc attagtatca
     1261 atttgcttac ctgtattcct ttacatcctc ctttttctcc ttcttgataa atgtatgtag
     1321 attgcgtata tagtttcgtc taccctatga acatattcca ttttgtaatt tcgtgtcgtt
     1381 tctattatga atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt
     1441 ttaagcaagg attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt
     1501 ggaaccacct aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc
     1561 aatggcctta ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa
     1621 gatagtggcg atagggttga ccttattctt tggcaaatct ggagcagaac cgtggcatgg
     1681 ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa
     1741 caaacccaag gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt
     1801 gctggtgatt ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga
     1861 atcaatcaat tgatgttgaa ccttcaatgt agggaattcg ttcttgatgg tttcctccac
     1921 agtttttctc cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg
     1981 caatggtggc tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc
     2041 tggaacggtg tattgttcac tatcccaagc gacaccatca ccatcgtctt cctttctctt
     2101 accaaagtaa atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa
     2161 ttgtggcttg attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa
     2221 gttggcgtac aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact
     2281 accggtaccc catttaggac cacccacagc acctaacaaa acggcatcag ccttcttgga
     2341 ggcttccagc gcctcatctg gaagtggaac acctgtagca tcgatagcag caccaccaat
     2401 taaatgattt tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac
     2461 cttaatggct tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt
     2521 cttaggggca gacattagaa tggtatatcc ttgaaatata tatatatatt gctgaaatgt
     2581 aaaaggtaag aaaagttaga aagtaagacg attgctaacc acctattgga aaaaacaata
     2641 ggtccttaaa taatattgtc aacttcaagt attgtgatgc aagcatttag tcatgaacgc
     2701 ttctctattc tatatgaaaa gccggttccg gcgctctcac ctttcctttt tctcccaatt
     2761 tttcagttga aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa aatttccagt
     2821 catcgaattt gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt gaggaaaaaa
     2881 ataatggttg ctaagagatt cgaactcttg catcttacga tactgagtat tcccacagtt
     2941 ccttaccact cttttgttac tctattgatc cagctcagca aaggcagtgt gatctaagat
     3001 tctatcttcg cgatgtagta aaactagcta gaccgagaaa gagactagaa atgcaaaagg
     3061 cacttctaca atggctgcca tcattattat ccgatgtgac gctgcagctt ctcaatgata
     3121 ttcgaatacg ctttgaggag atacagccta atatccgaca aactgtttta cagatttacg
     3181 atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa
     3241 cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg
     3301 tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg
     3361 catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg
     3421 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     3481 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     3541 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     3601 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     3661 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     3721 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     3781 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     3841 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     3901 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     3961 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     4021 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     4081 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     4141 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     4201 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     4261 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     4321 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg
     4381 aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga
     4441 aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg
     4501 cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg
     4561 tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta
     4621 cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct
     4681 ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca
     4741 tttcctttga tattggatca tatgcatagt accgagaaac tagtgcgaag tagtgatcag
     4801 gtattgctgt tatctgatga gtatacgttg tcctggccac ggcagaagca cgcttatcgc
     4861 tccaatttcc cacaacatta gtcaactccg ttaggccctt cattgaaaga aatgaggtca
     4921 tcaaatgtct tccaatgtga gattttgggc cattttttat agcaaagatt gaataaggcg
     4981 catttttctt caaagcttta ttgtacgatc tgactaagtt atcttttaat aattggtatt
     5041 cctgtttatt gcttgaagaa ttgccggtcc tatttactcg ttttaggact ggttcagaat
     5101 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5161 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5521 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     5581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5821 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     5881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6001 aaatctggag ccggtgagcg tggatctcgc ggtatcattg cagcactggg gccagatggt
     6061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6181 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6241 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6301 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6361 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6421 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6481 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6541 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6601 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6661 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6721 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6781 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6841 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6901 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6961 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7021 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7081 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     7141 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     7201 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     7261 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7321 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7381 aacgcgcgag gcagcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc
     7441 tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca
     7501 aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg
     7561 aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct
     7621 tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa
     7681 atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct
     7741 tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac tccaagcttt
     7801 gcaaagatgg ataaagcgga attaattccc gagcctccaa aaaagaagag aaaggtcgaa
     7861 ttgggtaccg ccgccaattt taatcaaagt gggaatattg ctgatagctc attgtccttc
     7921 actttcacta acagtagcaa cggtccgaac ctcataacaa ctcaaacaaa ttctcaagcg
     7981 ctttcacaac caattgcctc ctctaacgtt catgataact tcatgaataa tgaaatcacg
     8041 gctagtaaaa ttgatgatgg taataattca aaaccactgt cacctggttg gacggaccaa
     8101 actgcgtata acgcgtttgg aatcactaca gggatgttta ataccactac aatggatgat
     8161 gtatataact atctattcga tgatgaagat accccaccaa acccaaaaaa agagatctgg
     8221 aattcggatc ctcgagattt acctgagatg actcccgcag ttgcaatgac tcttagctta
     8281 gcagccaaca ccatgtgtga atcatcacct gtcgagatca ctcagctaaa gaacgttact
     8341 gatgcagctg acttgttatc tgattctgaa aatcaaagct tttgcaacgg agggactgaa
     8401 tgcactatgg aagatgtttc tgaactggaa gaggtaggtg aacaggattt gttgaaaact
     8461 ttatccgata cgagaagcgg gtcttccaat gtttttgatg aagacgatgt attgtctgtt
     8521 gtggaggata atagtgctgt cataagtgag ggcttgttag ttgttgatgc aggctctgaa
     8581 ttaagcttgt ctaatacagc tatggaaata gataacgggc gagttcttgc aaccgcgatt
     8641 atcgtaggcg aatcaagcat tgagcaggtt cccaccgcgg aagttcttat cgcgggtgta
     8701 aatcaggata ccaatacttc ggaggttgtc attagattgc
//
