LOCUS       sequence-360582-        8119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast-two-hybrid AD vector with 4XLexOp (activated by Lex-ER-B112).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8119)
  AUTHORS   Feuer E, Zimran G, Shpilman M, Mosquna A
  TITLE     A Modified Yeast Two-Hybrid Platform Enables Dynamic Control of 
            Expression Intensities to Unmask Properties of Protein-Protein 
            Interactions.
  JOURNAL   ACS Synth Biol. 2022 Jul 27. doi: 10.1021/acssynbio.2c00192.
  PUBMED    35895499
REFERENCE   2  (bases 1 to 8119)
  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: "ACS Synth 
            Biol. 2022 Jul 27. doi: 10.1021/acssynbio.2c00192."
COMMENT     Sequence Label: pACT-Lex
FEATURES             Location/Qualifiers
     source          1..8119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      112..299
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(167..191)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    complement(320..353)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(431..464)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(508..541)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(619..652)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1161..2255)
                     /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(2256..2661)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      3138..4480
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4101..4148)
                     /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)."
     promoter        4745..4849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4850..4918,4919..5710)
                     /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(5068..5087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5881..6469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6370..6389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6623..6640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6655..6795
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6721..6740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6881..6903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             7379..7399
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7415..7756
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             7769..7795
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7775..7801
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
ORIGIN
        1 ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct tgccagaaat ttacgaaaag
       61 atggaaaagg gtcaaatcgt tggtagatac gttgttgaca cttctaaata agcgaatttc
      121 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
      181 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
      241 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
      301 atgccggtcg agggacctaa taacttcgta tagcatacat tatacgaagt tatattaagg
      361 gttccggatc gcggccgctc gacctgcagc caagctagct tggctggacg taaactcctc
      421 ttcagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttattgaa
      481 tatgatcgga attggtcgag ggacctaata acttcgtata gcatacatta tacgaagtta
      541 tattaagggt tccggatcgc ggccgctcga cctgcagcca agctagcttg gctggacgta
      601 aactcctctt cagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg
      661 ttattgaata tgatcggaat tggtcgacta cgtcgttaag gccgtttctg acagagtaaa
      721 attcttgagg gaactttcac cattatggga aatggttcaa gaaggtattg acttaaactc
      781 catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt ttttagagaa
      841 aatcctccaa tatataaatt aggaatcata gtttcatgat tttctgttac acctaacttt
      901 ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag tgcaattctt tttccttatc
      961 acgttgagcc attagtatca atttgcttac ctgtattcct ttacatcctc ctttttctcc
     1021 ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga acatattcca
     1081 ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat gtacaaatat
     1141 cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc gacagcatca
     1201 ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc tgcatccaaa
     1261 acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa tgacaatttc
     1321 aacatcattg cagcagacaa gatagtggcg atagggttga ccttattctt tggcaaatct
     1381 ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag
     1441 gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc ttcatcggag
     1501 atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt tgggttctta
     1561 actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt aggaaattcg
     1621 ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa aacattagct
     1681 ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa agcggccatt
     1741 cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc gacaccatca
     1801 ccatcgtctt cctttctctt accaaagtaa atacctccca ctaattctct gacaacaacg
     1861 aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag agagtcggat
     1921 gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat ttttagtaaa
     1981 ccttgttcag gtctaacact acctgtaccc catttaggac cacccacagc acctaacaaa
     2041 acggcatcag ccttcttgga ggcttccagc gcctcatctg gaagtggaac acctgtagca
     2101 tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt ggaacgaaca
     2161 tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc aacgtggtca
     2221 cctggcaaaa cgacgatctt cttaggggca gacattagaa tggtatatcc ttgaaatata
     2281 tatatatatt gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc
     2341 acctattgga aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc
     2401 aagcatttag tcatgaacgc ttctctattc tatatgaaaa gccggttccg gcgctctcac
     2461 ctttcctttt tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa
     2521 attaacaaaa aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc
     2581 tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga
     2641 tacctgagta ttcccacagt tccttaccac tcttttgtta ctctattgat ccagctcagc
     2701 aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt aaaactagct agaccgagaa
     2761 agagactaga aatgcaaaag gcacttctac aatggctgcc atcattatta tccgatgtga
     2821 cgctgcagct tctcaatgat attcgaatac gctttgagga gatacagcct aatatccgac
     2881 aaactgtttt acagatttac gatcgtactt gttacccatc attgaatttt gaacatccga
     2941 acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa tatatagtct
     3001 agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt gcatctattg
     3061 cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat cttgcacttc
     3121 aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg
     3181 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3241 acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg
     3301 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3361 atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa
     3421 aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct
     3481 ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta
     3541 gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc
     3601 gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga
     3661 ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat
     3721 gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg
     3781 tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta
     3841 caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt
     3901 tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga
     3961 gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt
     4021 agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt
     4081 ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata
     4141 ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca
     4201 tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg
     4261 agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt
     4321 aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta
     4381 tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag
     4441 tctcatcctt caatgctatc atttcctttg atattggatc atatgcatag taccgagaaa
     4501 ctagtgcgaa gtagtgatca ggtattgctg ttatctgatg agtatacgtt gtcctggcca
     4561 cggcagaagc acgcttatcg ctccaatttc ccacaacatt agtcaactcc gttaggccct
     4621 tcattgaaag aaatgaggtc atcaaatgtc ttccaatgtg agattttggg ccatttttta
     4681 tagcaaagat tgaataaggc gcatttttct tcaaagctca ggtggcactt ttcggggaaa
     4741 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4801 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4861 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4921 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4981 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5041 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     5101 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5161 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5221 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5281 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5341 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     5401 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5461 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5521 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5581 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5641 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5701 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5761 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6001 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6541 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6601 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     6661 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     6721 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     6781 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6901 aggttttcac cgtcatcacc gaaacgcttc gatactaacg ccgccatcca gtgtttaaac
     6961 gaactagtgc ggccgcaata atatataaac ctgtataata taaccttgaa gactatattt
     7021 cttttcgagc tccctaggtg ctgtatatac tcacagcata actgtatata cacccagggt
     7081 ctaggtgctg tatatactca cagcataact gtatatacac ccagggtcta ggtgctgtat
     7141 atactcacag cataactgta tatacaccca gggtctaggt gctgtatata ctcacagcat
     7201 aactgtatat acacccaggg tctagagcat gtgctctgta tgtatataaa actcttgttt
     7261 tcttcttttc tcttctttcc ttatacatta ggacctttgc agcataaatt actatacttc
     7321 tatactagtg gatcccccgg gctgcaggat ggataaagcg gaattaattc ccgagcctcc
     7381 aaaaaagaag agaaaggtcg aattgggtac cgccgccaat tttaatcaaa gtgggaatat
     7441 tgctgatagc tcattgtcct tcactttcac taacagtagc aacggtccga acctcataac
     7501 aactcaaaca aattctcaag cgctttcaca accaattgcc tcctctaacg ttcatgataa
     7561 cttcatgaat aatgaaatca cggctagtaa aattgatgat ggtaataatt caaaaccact
     7621 gtcacctggt tggacggacc aaactgcgta taacgcgttt ggaatcacta cagggatgtt
     7681 taataccact acaatggatg atgtatataa ctatctattc gatgatgaag ataccccacc
     7741 aaacccaaaa aaagagatct ctatggctta cccatacgat gttccagatt acgctagctt
     7801 gggtggtcat atggccatgg aggccccggg gatccgaatt cgagctcgag agatctatga
     7861 atcgtagata ctgaaaaacc ccgcaagttc acttcaactg tgcatcgtgc accatctcaa
     7921 tttctttcat ttatacatcg ttttgccttc ttttatgtaa ctatactcct ctaagtttca
     7981 atcttggcca tgtaacctct gatctataga attttttaaa tgactagaat taatgcccat
     8041 cttttttttg gacctaaatt cttcatgaaa atatattacg agggcttatt cagaagcttt
     8101 ggacttcttc gccagaggt
//
