LOCUS       sequence-484548-        9528 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Yeast two hybrid vector expressing AD-HAB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9528)
  AUTHORS   Park SY, Fung P, Nishimura N, Jensen DR, Fujii H, Zhao Y, Lumba S, 
            Santiago J, Rodrigues A, Chow TF, Alfred SE, Bonetta D, Finkelstein 
            R, Provart NJ, Desveaux D, Rodriguez PL, McCourt P, Zhu JK, 
            Schroeder JI, Volkman BF, Cutler SR
  TITLE     Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL 
            family of START proteins.
  JOURNAL   Science. 2009 May 22;324(5930):1068-71. doi: 
            10.1126/science.1173041. Epub 2009 Apr 30.
  PUBMED    19407142
REFERENCE   2  (bases 1 to 9528)
  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: 
            "10.1126/science.1173041"; journalName: "Science"; date: 
            "2009-05-22- 22"; volume: "324"; issue: "5930"; pages: "1068-71"
COMMENT     Sequence Label: pACT-HAB1
FEATURES             Location/Qualifiers
     source          1..9528
                     /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(1417..2511)
                     /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(2512..2916)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      3393..4735
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4356..4403)
                     /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(5093..5111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5179..5283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5284..5352,5353..6144)
                     /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(5502..5521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6315..6903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6804..6823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7057..7074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7089..7229
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7155..7174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7315..7337
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        7366..7767
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             7813..7833
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7849..8190
                     /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 aagctcaaag 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 agagaaaatc ctccaatata taaattagga atcatagttt catgattttc
     1141 tgttacacct aactttttgt gtggtgccct cctccttgtc aatattaatg ttaaagtgca
     1201 attctttttc cttatcacgt tgagccatta gtatcaattt gcttacctgt attcctttac
     1261 atcctccttt ttctccttct tgataaatgt atgtagattg cgtatatagt ttcgtctacc
     1321 ctatgaacat attccatttt gtaatttcgt gtcgtttcta ttatgaattt catttataaa
     1381 gtttatgtac aaatatcata aaaaaagaga atctttttaa gcaaggattt tcttaacttc
     1441 ttcggcgaca gcatcaccga cttcggtggt actgttggaa ccacctaaat caccagttct
     1501 gatacctgca tccaaaacct ttttaactgc atcttcaatg gccttacctt cttcaggcaa
     1561 gttcaatgac aatttcaaca tcattgcagc agacaagata gtggcgatag ggttgacctt
     1621 attctttggc aaatctggag cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt
     1681 cttgtctggc aaagaggcca aggacgcaga tggcaacaaa cccaaggaac ctgggataac
     1741 ggaggcttca tcggagatga tatcaccaaa catgttgctg gtgattataa taccatttag
     1801 gtgggttggg ttcttaacta ggatcatggc ggcagaatca atcaattgat gttgaacctt
     1861 caatgtaggg aattcgttct tgatggtttc ctccacagtt tttctccata atcttgaaga
     1921 ggccaaaaca ttagctttat ccaaggacca aataggcaat ggtggctcat gttgtagggc
     1981 catgaaagcg gccattcttg tgattctttg cacttctgga acggtgtatt gttcactatc
     2041 ccaagcgaca ccatcaccat cgtcttcctt tctcttacca aagtaaatac ctcccactaa
     2101 ttctctgaca acaacgaagt cagtaccttt agcaaattgt ggcttgattg gagataagtc
     2161 taaaagagag tcggatgcaa agttacatgg tcttaagttg gcgtacaatt gaagttcttt
     2221 acggattttt agtaaacctt gttcaggtct aacactaccg gtaccccatt taggaccacc
     2281 cacagcacct aacaaaacgg catcagcctt cttggaggct tccagcgcct catctggaag
     2341 tggaacacct gtagcatcga tagcagcacc accaattaaa tgattttcga aatcgaactt
     2401 gacattggaa cgaacatcag aaatagcttt aagaacctta atggcttcgg ctgtgatttc
     2461 ttgaccaacg tggtcacctg gcaaaacgac gatcttctta ggggcagaca ttagaatggt
     2521 atatccttga aatatatata tatattgctg aaatgtaaaa ggtaagaaaa gttagaaagt
     2581 aagacgattg ctaaccacct attggaaaaa acaataggtc cttaaataat attgtcaact
     2641 tcaagtattg tgatgcaagc atttagtcat gaacgcttct ctattctata tgaaaagccg
     2701 gttccggcgc tctcaccttt cctttttctc ccaatttttc agttgaaaaa ggtatatgcg
     2761 tcaggcgacc tctgaaatta acaaaaaatt tccagtcatc gaatttgatt ctgtgcgata
     2821 gcgcccctgt gtgttctcgt tatgttgagg aaaaaaataa tggttgctaa gagattcgaa
     2881 ctcttgcatc ttacgatact gagtattccc acagttcctt accactcttt tgttactcta
     2941 ttgatccagc tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac
     3001 tagctagacc gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat
     3061 tattatccga tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac
     3121 agcctaatat ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga
     3181 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     3241 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     3301 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     3361 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     3421 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3481 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     3541 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     3601 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     3661 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     3721 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     3781 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     3841 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     3901 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     3961 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     4021 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     4081 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     4141 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     4201 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     4261 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     4321 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     4381 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     4441 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     4501 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     4561 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     4621 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     4681 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatatg
     4741 catagtaccg agaaactagt gcgaagtagt gatcaggtat tgctgttatc tgatgagtat
     4801 acgttgtcct ggccacggca gaagcacgct tatcgctcca atttcccaca acattagtca
     4861 actccgttag gcccttcatt gaaagaaatg aggtcatcaa atgtcttcca atgtgagatt
     4921 ttgggccatt ttttatagca aagattgaat aaggcgcatt tttcttcaaa gctttattgt
     4981 acgatctgac taagttatct tttaataatt ggtattcctg tttattgctt gaagaattgc
     5041 cggtcctatt tactcgtttt aggactggtt cagaattctt gaagacgaaa gggcctcgtg
     5101 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5161 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5521 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5821 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     6001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6421 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     6481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6961 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7021 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7081 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     7141 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     7201 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     7261 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7321 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag caacttcttt
     7381 tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca atgacaaaaa
     7441 aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt
     7501 gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc cttcattcac
     7561 gcacactact ctctaatgag caacggtata cggccttcct tccagttact tgaatttgaa
     7621 ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt attaatcttt
     7681 tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg cacaatattt
     7741 caagctatac caagcataca atcaactcca agctttgcaa agatggataa agcggaatta
     7801 attcccgagc ctccaaaaaa gaagagaaag gtcgaattgg gtaccgccgc caattttaat
     7861 caaagtggga atattgctga tagctcattg tccttcactt tcactaacag tagcaacggt
     7921 ccgaacctca taacaactca aacaaattct caagcgcttt cacaaccaat tgcctcctct
     7981 aacgttcatg ataacttcat gaataatgaa atcacggcta gtaaaattga tgatggtaat
     8041 aattcaaaac cactgtcacc tggttggacg gaccaaactg cgtataacgc gtttggaatc
     8101 actacaggga tgtttaatac cactacaatg gatgatgtat ataactatct attcgatgat
     8161 gaagataccc caccaaaccc aaaaaaagag atctggaatt cggatcctcg agatttacct
     8221 gagatgactc ccgcagttgc aatgactctt agcttagcag ccaacaccat gtgtgaatca
     8281 tcacctgtcg agatcactca gctaaagaac gttactgatg cagctgactt gttatctgat
     8341 tctgaaaatc aaagcttttg caacggaggg actgaatgca ctatggaaga tgtttctgaa
     8401 ctggaagagg taggtgaaca ggatttgttg aaaactttat ccgatacgag aagcgggtct
     8461 tccaatgttt ttgatgaaga cgatgtattg tctgttgtgg aggataatag tgctgtcata
     8521 agtgagggct tgttagttgt tgatgcaggc tctgaattaa gcttgtctaa tacagctatg
     8581 gaaatagata acgggcgagt tcttgcaacc gcgattatcg taggcgaatc aagcattgag
     8641 caggttccca ccgcggaagt tcttatcgcg ggtgtaaatc aggataccaa tacttcggag
     8701 gttgtcatta gattgccaga tgaaaatagt aatcatctgg tgaaagggag aagtgtttat
     8761 gaactagatt gtataccgct ttggggcacg gtttccattc aagggaatag atctgagatg
     8821 gaggatgctt ttgccgtgtc acctcatttt ctgaaactac ccatcaaaat gcttatgggg
     8881 gaccatgagg gtatgagtcc aagcctcaca cacctcaccg gtcatttttt cggtgtttat
     8941 gatggtcatg gaggccataa ggttgctgac tattgccgag atagactcca ttttgctttg
     9001 gctgaagaaa tagaacgtat aaaagacgaa ttatgcaaga ggaatacagg ggagggtagg
     9061 caggtgcagt gggataaagt cttcacgagt tgttttctaa ctgtcgatgg tgagattgaa
     9121 ggaaaaattg gtagagccgt tgttggttct tctgataagg ttcttgaggc tgttgcgtct
     9181 gagactgtag gatcaactgc tgttgttgcc ttggtttgct catcacatat agtagtttct
     9241 aactgcggtg attcgagggc ggttttattc cgtggcaaag aagccatgcc cttgtcagtt
     9301 gatcacaaac cagatagaga ggatgaatat gcaagaatag aaaatgctgg aggcaaagtt
     9361 atacaatggc aaggcgcacg tgtttttggt gttctcgcca tgtctaggtc catcggtgac
     9421 agatatctga agccatatgt gatcccagaa ccggaagtga cattcatgcc tcggtcaaga
     9481 gaagacgagt gtctcatact agccagtgac ggtctttggg atgtaatg
//
