LOCUS       sequence-77667-e        7036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bi-directional expression vector for yeast, constitutively active, 
            LEU2 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  AUTHORS   Miller CA 3rd, Martinat MA, Hyman LE
  TITLE     Assessment of aryl hydrocarbon receptor complex interactions using 
            pBEVY plasmids: expressionvectors with bi-directional promoters for 
            use in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 1998 Aug 1;26(15):3577-83.
  PUBMED    9671822
REFERENCE   2  (bases 1 to 7036)
  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: "Nucleic 
            Acids Res."; date: "1998-08-1"; volume: "26(15)"; pages: "3577-83"
COMMENT     Sequence Label: pBEVY-L
FEATURES             Location/Qualifiers
     source          1..7036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(267..454)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     375..399
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(623..1268)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(708..731)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     promoter        1277..1673
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     terminator      1712..2011
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
     rep_origin      2036..3382
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3520..4614)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4626..5024)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5044..5062)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5130..5234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5235..5303,5304..6095)
                     /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(5453..5472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6266..6854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6755..6774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7008..7025
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctttt
      241 catagggtag gggaatttcg ggcatgccgg tagaggtgtg gtcaataaga gcgacctcat
      301 gctatacctg agaaagcaac ctgacctaca ggaaagagtt actcaagaac aagaattttc
      361 gttttaaaac ctaagagtca ctttaaaatt tgtatacact tatttttttt ataacttatt
      421 taataataaa aatcataaat cataagaaat tcgcttattt agaagtgtca acaacgtatc
      481 taccaacgat ttgacccttt tccatctttt cgtaaatttc tggcaaggta gacaagccga
      541 caaccttgat tggagacttg accaaacctc tggcgaagaa gtccaaagct tgcatgcctg
      601 caggtcgact ctagaggatc cgtcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa
      661 agactaacta taaaagtaga atttaagaag tttaagaaat agatttacag aattacaatc
      721 aatacctacc gttctttata tacttattag tcaagtaggg gaataatttc agggaactgg
      781 tttcaacctt ttttttcagc tttttccaaa tcagagagag cagaaggtaa tagaaggtgt
      841 aagaaaatga gatagataca tgcgtgggtc aattgccttg tgtcatcatt tactccaggc
      901 aggttgcatc actccattga ggttgtgccc gttttttgcc tgtttgtgcc cctgttctct
      961 gtagttgcgc taagagaatg gacctatgaa ctgatggttg gtgaagaaaa caatattttg
     1021 gtgctgggat tctttttttt tctggatgcc agcttaaaaa gcgggctcca ttatatttag
     1081 tggatgccag gaataaactg ttcacccaga cacctacgat gttatatatt ctgtgtaacc
     1141 cgccccctat tttgggcatg tacgggttac agcagaatta aaaggctaat tttttgacta
     1201 aataaagtta ggaaaatcac tactattaat tatttacgta ttctttgaaa tggcagtatt
     1261 gataatgata aactcccgga ttcttttctt tttttttctt ttctctctcc cccgttgttg
     1321 tctcaccata tccgcaatga caaaaaaaat gatggaagac actaaaggaa aaaattaacg
     1381 acaaagacag caccaacaga tgtcgttgtt ccagagctga tgaggggtat cttcgaacac
     1441 acgaaacttt ttccttcctt cattcacgca cactactctc taatgagcaa cggtatacgg
     1501 ccttccttcc agttacttga atttgaaata aaaaaagttt gccgctttgc tatcaagtat
     1561 aaatagacct gcaattatta atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc
     1621 cttgtttctt tttctgcaca atatttcaag ctataccaag catacaatca actccccggg
     1681 taccgagctc gaattcgaca cttctaaata agcggatctc ctatgccttc acgatttata
     1741 gtttccatta tcaagtatgc ctatattagt atatagcatc tttagatgac agtgttcgaa
     1801 gtttcacgaa taaaagataa tattctactt tttgctccca ccgcgtttgc tagcacgagt
     1861 gaacaccatc cctcgcctgt gagttgtacc cattcctcta aactgtagac atggtagctt
     1921 cagcagtgtt cgttatgtac ggcatcctcc aacaaacagt cggttatagt ttgtcctgct
     1981 cctctgaatc gtctccctcg atatttctca ttttccttcg catgccctcg agcggaacga
     2041 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2101 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2161 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     2221 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     2281 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     2341 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     2401 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     2461 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     2521 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     2581 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     2641 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     2701 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     2761 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     2821 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     2881 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     2941 cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga
     3001 agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc aaagcgtttc
     3061 cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc actgttcacg
     3121 tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac ggcatagtgc
     3181 gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa aggtagtcta
     3241 gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct tcagcactac
     3301 cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc ttcaatgcta
     3361 tcatttcctt tgatattgga tcgatccgat gataagctgt caaacatgag aattaattct
     3421 accctatgaa catattccat tttgtaattt cgtgtcgttt ctattatgaa tttcatttat
     3481 aaagtttatg tacaaatatc ataaaaaaag agaatctttt taagcaagga ttttcttaac
     3541 ttcttcggcg acagcatcac cgacttcggt ggtactgttg gaaccaccta aatcaccagt
     3601 tctgatacct gcatccaaaa cctttttaac tgcatcttca atggccttac cttcttcagg
     3661 caagttcaat gacaatttca acatcattgc agcagacaag atagtggcga tagggtcaac
     3721 cttattcttt ggcaaatctg gagcagaacc gtggcatggt tcgtacaaac caaatgcggt
     3781 gttcttgtct ggcaaagagg ccaaggacgc agatggcaac aaacccaagg aacctgggat
     3841 aacggaggct tcatcggaga tgatatcacc aaacatgttg ctggtgatta taataccatt
     3901 taggtgggtt gggttcttaa ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac
     3961 cttcaatgta ggaaattcgt tcttgatggt ttcctccaca gtttttctcc ataatcttga
     4021 agaggccaaa acattagctt tatccaagga ccaaataggc aatggtggct catgttgtag
     4081 ggccatgaaa gcggccattc ttgtgattct ttgcacttct ggaacggtgt attgttcact
     4141 atcccaagcg acaccatcac catcgtcttc ctttctctta ccaaagtaaa tacctcccac
     4201 taattctctg acaacaacga agtcagtacc tttagcaaat tgtggcttga ttggagataa
     4261 gtctaaaaga gagtcggatg caaagttaca tggtcttaag ttggcgtaca attgaagttc
     4321 tttacggatt tttagtaaac cttgttcagg tctaacacta cctgtacccc atttaggacc
     4381 acccacagca cctaacaaaa cggcatcaac cttcttggag gcttccagcg cctcatctgg
     4441 aagtgggaca cctgtagcat cgatagcagc accaccaatt aaatgatttt cgaaatcgaa
     4501 cttgacattg gaacgaacat cagaaatagc tttaagaacc ttaatggctt cggctgtgat
     4561 ttcttgacca acgtggtcac ctggcaaaac gacgatcttc ttaggggcag acataggggc
     4621 agacattaga atgtatatcc ttgaaatata tatatatatt gctgaaatgt aaaaggtaag
     4681 aaaagttaga ataagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata
     4741 atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta
     4801 tatgaaaagc cggttccggc ctctcacctt tcctttttct cccaattttt cagttgaaaa
     4861 aggtatatgc gcggcgacct ctgaaattaa caaaaaattt ccagtcatcg aatttgtctg
     4921 tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag
     4981 attcgaactc ttgcatctta cgatacctga gtattcccac agttaattct tgaagacgaa
     5041 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5101 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5161 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5221 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5281 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5341 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5401 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5461 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5521 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5581 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5641 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5701 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5761 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5821 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5881 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5941 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6001 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6061 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6121 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6181 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6241 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6301 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6361 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6721 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6781 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6841 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6901 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6961 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7021 gcgaggaagc ggaaga
//
