LOCUS       sequence-254980-        8176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8176)
  AUTHORS   Worseck JM, Grossmann A, Weimann M, Hegele A, Stelzl U
  TITLE     A stringent yeast two-hybrid matrix screening approach for 
            protein-protein interaction discovery.
  JOURNAL   Methods Mol Biol. 2012;812:63-87. doi: 10.1007/978-1-61779-455-1_4.
  PUBMED    22218854
REFERENCE   2  (bases 1 to 8176)
  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: "Methods Mol
            Biol."; date: "2012"; volume: "812:63-87. doi"; pages: " 
            10.1007/978-1-61779-455-1_4"
COMMENT     Sequence Label: pBTM116-D9
FEATURES             Location/Qualifiers
     source          1..8176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             join(433..684,685..1038)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     976..997
                     /label=LexA
                     /note="LexA DNA binding domain, forward primer"
     protein_bind    1057..1181
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1355..1457
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1458..2117
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1524..1543)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2459..2764
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTADMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2805..2929)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2974..3161
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3029..3053)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3214..3888)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3889..3990)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4004..4020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4004..4020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4017..4039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4028..4044
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4052..4058,4059..4076,4077..4082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4097..4118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4235..4252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4406..4994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4486..4505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(5274..5378)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5491..5496,5497..5513,5514..5519)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             5567..6757
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(5919..5938)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6854..6876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6976..6995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7012..8176
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
      121 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
      181 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
      241 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaatta
      421 attccgggcg gaatgaaagc gttaacggcc aggcaacaag aggtgtttga tctcatccgt
      481 gatcacatca gccagacagg tatgccgccg acgcgtgcgg aaatcgcgca gcgtttgggg
      541 ttccgttccc caaacgcggc tgaagaacat ctgaaggcgc tggcacgcaa aggcgttatt
      601 gaaattgttt ccggcgcatc acgcgggatt cgtctgttgc aggaagagga agaagggttg
      661 ccgctggtag gtcgtgtggc tgccggtgaa ccacttctgg cgcaacagca tattgaaggt
      721 cattatcagg tcgatccttc cttattcaag ccgaatgctg atttcctgct gcgcgtcagc
      781 gggatgtcga tgaaagatat cggcattatg gatggtgact tgctggcagt gcataaaact
      841 caggatgtac gtaacggtca ggtcgttgtc gcacgtattg atgacgaggt taccgttaag
      901 cgcctgaaaa aacagggcaa taaagtcgaa ctgttgccag aaaatagcga gtttaaacca
      961 attgtcgtag atcttcgtca gcagagcttc accattgaag ggctggcggt tggggttatt
     1021 cgcaacggcg actggctgga attcatgtcg aatcaaacaa gtttgtacaa aaaagctgaa
     1081 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     1141 actacataat actgtaaaac acaacatatc cagtcactat ggcggccgcg ggtgatgctg
     1201 ccaacttagc ggccgctaag ttggcagcat cacccgacgc actttgcgcc gaataaatac
     1261 ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga
     1321 agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact
     1381 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     1441 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1501 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1561 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1621 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     1681 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1741 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1801 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     1861 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     1921 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     1981 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2041 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2101 ggcagggcgg ggcgtaatct agaggatccg gcttactaaa agccagataa cagtatgcgt
     2161 atttgcgcgc tgattttcgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     2221 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     2281 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     2341 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     2401 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     2461 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     2521 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     2581 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     2641 gcgcatgatg accgccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     2701 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     2761 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     2821 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     2881 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt cgattcgacg
     2941 gtaccgcggc cgctgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt
     3001 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     3061 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     3121 gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccggc aagtgcacaa
     3181 acaatactta aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt
     3241 tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc
     3301 aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa
     3361 cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca
     3421 atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg
     3481 tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat
     3541 aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac
     3601 gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa
     3661 cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta caagacttga
     3721 aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc
     3781 cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca
     3841 aactttcacc aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc
     3901 ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg
     3961 gccctctcct tttctttttt cgaccgaatt aattcgtaat catgtcatag ctgtttcctg
     4021 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4081 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5161 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5221 ccatagttgc ctgactcggg aataagggcg acacggaaat gttgaatact catactcttc
     5281 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5341 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5401 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5461 aggccctttc gtcttcaaga attctcatgt ttgacagctt atcatcgata agctttaatg
     5521 cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa
     5581 tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat
     5641 gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg ccagtcacta
     5701 tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc
     5761 actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat
     5821 cgactacgcg atcatggcga ccacacccgt cctgtggatc ctctacgccg gacgcatcgt
     5881 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     5941 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     6001 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     6061 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     6121 taagggagag cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     6181 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     6241 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     6301 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     6361 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     6421 ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc
     6481 cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc
     6541 caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag
     6601 cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac
     6661 atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt
     6721 gcgtcgcggt gcatggagcc gggccacctc gacctgaccc ctaagaaacc attattatca
     6781 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6841 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6901 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6961 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aacgcattta
     7021 agcataaaca cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga
     7081 tataggtgcg acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc
     7141 gctcgttttc ggaaacgctt tgaagttcct attccgaagt tcctattctc tagctagaaa
     7201 gtataggaac ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa
     7261 aaccaaaaac gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa
     7321 acattgctca aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac
     7381 ccatccacct ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt
     7441 atctctagta ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc
     7501 gaaaacaata cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga
     7561 aaccgttcat aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa
     7621 agtatgcgca atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat
     7681 gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg
     7741 gaagagaaaa tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa
     7801 agttatcaag agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct
     7861 ttgttagaaa aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt
     7921 ctttgttggt aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     7981 ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac
     8041 agattcttcg ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc
     8101 tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa
     8161 gcacagatgc ttcgtt
//
