LOCUS       sequence-393055-        8421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast two-hybrid ; Gal4 activation domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  AUTHORS   Uetz P, Giot L, Cagney G, Mansfield TA, Judson RS, Knight JR, 
            Lockshon D, Narayan V, Srinivasan M, Pochart P, Qureshi-Emili A, Li 
            Y, Godwin B, Conover D, Kalbfleisch T, Vijayadamodar G, Yang M, 
            Johnston M, Fields S, Rothberg JM
  TITLE     A comprehensive analysis of protein-protein interactions in 
            Saccharomyces cerevisiae.
  JOURNAL   Nature. 2000 Feb 10;403(6770):623-7. doi: 10.1038/35001009.
  PUBMED    10688190
REFERENCE   2  (bases 1 to 8421)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2000-02-10- 10"; volume: "403"; issue: "6770"; 
            pages: "623-7"
COMMENT     Sequence Label: pOAD
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      408..595
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(463..487)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1425..2587
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      1750..2587
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(2949..4043)
                     /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(4044..4449)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4469..4487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4555..4659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4660..4728,4729..5520)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4878..4897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6433..6450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7268..7972
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             8018..8038
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8054..8395
                     /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 aattcccggg gatccgtcga cctgcagaga tctatgaatc gtagatactg aaaaaccccg
       61 caagttcact tcaactgtgc atcgtgcacc atctcaattt ctttcattta tacatcgttt
      121 tgccttcttt tatgtaacta tactcctcta agtttcaatc ttggccatgt aacctctgat
      181 ctatagaatt ttttaaatga ctagaattaa tgcccatctt ttttttggac ctaaattctt
      241 catgaaaata tattacgagg gcttattcag aagctttgga cttcttcgcc agaggtttgg
      301 tcaagtctcc aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg
      361 aaaagggtca aatcgttggt agatacgttg ttgacacttc taaataagcg aatttcttat
      421 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
      481 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
      541 ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct accggcatgc
      601 cgagcaaatg cctgcaaatc gctccccatt tcacccaatt gtagatatgc taactccagc
      661 aatgagttga tgaatctcgg tgtgtatttt atgtcctcag aggacaacac ctgttgtaat
      721 cgttcttcca cacggatcct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      781 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
      841 gtgcggtatt tcacaccgca taatcgctgg gccattctca tgaagaatat cttgaattta
      901 ttgtcatatt actagttggt gtggaagtcc taatatcggt gatcaatata gtggttgaca
      961 tgctggctag tcaacattga gccttttgat catgcaaata tattacggta ttttacaatc
     1021 aaatatcaaa cttaactatt gactttataa cttatttagg tggtaacatt cttataaaaa
     1081 agaaaaaaat tactgcaaaa cagtactagc ttttaacttg tatcctaggt tatctatgct
     1141 gtctcaccat agagaatatt acctatttca gaatgtatgt ccatgattcg ccgggtaaat
     1201 acatataata cacaaatctg gcttaataaa gtctataata tatctcataa agaagtgcta
     1261 aattggctag tgctatatat ttttaagaaa atttcttttg actaagtcca tatcgacttt
     1321 gtaaaagttc acattagcat acatatatta cacgagccag aaatagtaac ttttgcctaa
     1381 atcacaaatt gcaaaattta attgcttgca aaaggtcaca tgcttataat caactttttt
     1441 aaaaatttaa aatacttttt tattttttat ttttaaacat aaatgaaata atttatttat
     1501 tgtttatgat taccgaaaca taaaacctgc tcaagaaaaa gaaactgttt tgtccttgga
     1561 aaaaaagcac tacctaggag cggccaaaat gccgaggctt tcatagctta aactctttac
     1621 agaaaatagg cattatagat cagttcgagt tttcttattc ttccttccgg ttttatcgtc
     1681 acagttttac agtaaataag tatcacctct tagagttcga tgataagctg tcaaacatga
     1741 gaattaattc cacatgttaa aatagtgaag gagcatgttc ggcacacagt ggaccgaacg
     1801 tggggtaagt gcactagggt ccggttaaac ggatctcgca ttgatgaggc aacgctaatt
     1861 atcaacatat agattgttat ctatctgcat gaacacgaaa tctttacttg acgacttgag
     1921 gctgatggtg tttatgcaaa gaaaccactg tgtttaatat gtgtcactgt ttgatattac
     1981 tgtcagcgta gaagataata gtaaaagcgg ttaataagtg tatttgagat aagtgtgata
     2041 aagtttttac agcgaaaaga cgataaatac aagaaaatga ttacgaggat acggagagag
     2101 gtatgtacat gtgtatttat atactaagct gccggcggtt gtttgcaaga ccgagaaaag
     2161 gctagcaaga atcgggtcat tgtagcgtat gcgcctgtga acattctctt caacaagttt
     2221 gattccattg cggtgaaatg gtaaaagtca accccctgcg atgtatattt tcctgtacaa
     2281 tcaatcaaaa agccaaatga tttagcatta tctttacatc ttgttatttt acagatttta
     2341 tgtttagatc ttttatgctt gcttttcaaa aggcttgcag gcaagtgcac aaacaatact
     2401 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt
     2461 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc
     2521 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat
     2581 gtaagctctc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa
     2641 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg
     2701 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca
     2761 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatcgatg
     2821 ataagctgtc aaacatgaga attaattcta ccctatgaac atattccatt ttgtaatttc
     2881 gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca taaaaaaaga
     2941 gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttcggtg
     3001 gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact
     3061 gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa catcattgca
     3121 gcagacaaga tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg
     3181 tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca
     3241 gatggcaaca aacccaagga acctgggata acggaggctt catcggagat gatatcacca
     3301 aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg
     3361 gcggcagaat caatcaattg atgttgaacc ttcaatgtag gaaattcgtt cttgatggtt
     3421 tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac
     3481 caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt
     3541 tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc
     3601 tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct
     3661 ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat
     3721 ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt
     3781 ctaacactac ctgtacccca tttaggacca cccacagcac ctaacaaaac ggcatcagcc
     3841 ttcttggagg cttccagcgc ctcatctgga agtggaacac ctgtagcatc gatagcagca
     3901 ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct
     3961 ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg
     4021 acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata tatatattgc
     4081 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa
     4141 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
     4201 atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct ttcctttttc
     4261 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa
     4321 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga
     4381 ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt
     4441 cccacagtta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4501 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4861 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5521 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6421 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6481 cctctccccg cgcgttggcc gattcattaa tgcaggatcc gggatcgaag aaatgatggt
     6541 aaatgaaata ggaaatcaag gagcatgaag gcaaaagaca aatataaggg tcgaacgaaa
     6601 aataaagtga aaagtgttga tatgatgtat ttggctttgc ggcgccgaaa aaacgagttt
     6661 acgcaattgc acaatcatgc tgactctgtg gcggacccgc gctcttgccg gcccggcgat
     6721 aacgctgggc gtgaggctgt gcccggcgga gttttttgcg cctgcatttt ccaaggttta
     6781 ccctgcgcta aggggcgaga ttggagaagc aataagaatg ccggttgggg ttgcgatgat
     6841 gacgaccacg acaactggtg tcattattta agttgccgaa agaacctgag tgcatttgca
     6901 acatgagtat actagaagaa tgagccaaga cttgcgagac gcgagtttgc cggtggtgcg
     6961 aacaatagag cgaccatgac cttgaaggtg agacgcgcat aaccgctaga gtactttgaa
     7021 gaggaaacag caatagggtt gctaccagta taaatagaca ggtacataca acactggaaa
     7081 tggttgtctg tttgagtacg ctttcaattc atttgggtgt gcactttatt atgttacaat
     7141 atggaaggga actttacact tctcctatgc acatatatta attaaagtcc aatgctagta
     7201 gagaaggggg gtaacacccc tccgcgctct tttccgattt ttttctaaac cgtggaatat
     7261 ttcggatatc cttttgttgt ttccgggtgt acaatatgga cttcctcttt tctggcaacc
     7321 aaacccatac atcgggattc ctataatacc ttcgttggtc tccctaacat gtaggtggcg
     7381 gaggggagat atacaataga acagatacca gacaagacat aatgggctaa acaagactac
     7441 accaattaca ctgcctcatt gatggtggta cataacgaac taatactgta gccctagact
     7501 tgatagccat catcatatcg aagtttcact accctttttc catttgccat ctattgaagt
     7561 aataataggc gcatgcaact tcttttcttt ttttttcttt tctctctccc ccgttgttgt
     7621 ctcaccatat ccgcaatgac aaaaaaatga tggaagacac taaaggaaaa aattaacgac
     7681 aaagacagca ccaacagatg tcgttgttcc agagctgatg aggggtatct cgaagcacac
     7741 gaaacttttt ccttccttca ttcacgcaca ctactctcta atgagcaacg gtatacggcc
     7801 ttccttccag ttacttgaat ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata
     7861 aatagacctg caattattaa tcttttgttt cctcgtcatt gttctcgttc cctttcttcc
     7921 ttgtttcttt ttctgcacaa tatttcaagc tataccaagc atacaatcaa ctccaagctt
     7981 tgcaaagatg gataaagcgg aattaattcc cgagcctcca aaaaagaaga gaaaggtcga
     8041 attgggtacc gccgccaatt ttaatcaaag tgggaatatt gctgatagct cattgtcctt
     8101 cactttcact aacagtagca acggtccgaa cctcataaca actcaaacaa attctcaagc
     8161 gctttcacaa ccaattgcct cctctaacgt tcatgataac ttcatgaata atgaaatcac
     8221 ggctagtaaa attgatgatg gtaataattc aaaaccactg tcacctggtt ggacggacca
     8281 aactgcgtat aacgcgtttg gaatcactac agggatgttt aataccacta caatggatga
     8341 tgtatataac tatctattcg atgatgaaga taccccacca aacccaaaaa aagagatcga
     8401 attccagctg accaccatgg c
//
