LOCUS       sequence-8858-ex        8001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Wang H, Heinz ME, Crosby SD, Johnston M, Mitra RD
  TITLE     'Calling Cards' method for high-throughput identification of targets
            of yeast DNA-binding proteins.
  JOURNAL   Nat Protoc. 2008;3(10):1569-77. doi: 10.1038/nprot.2008.148.
  PUBMED    18802438
REFERENCE   2  (bases 1 to 8001)
  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: "Nat 
            Protoc."; date: "2008"; volume: "3(10):1569-77. doi"; pages: " 
            10.1038/nprot.2008.148"
COMMENT     Sequence Label: pBM4607
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             434..874
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1694..1723
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1766..1795
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2190..2377
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2245..2269)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    3209..4371
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3534..4371
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(4210..4884)
                     /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(4885..4986)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5006..5024)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5092..5196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5197..5265,5266..6057)
                     /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(5415..5434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6228..6816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6717..6736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6970..6987
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc atatcgaagt
       61 ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat gcaacttctt
      121 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
      181 aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt
      241 tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgaactat ctattcgatg atgaagatac
      901 cccaccaaac ccaaaaaaag agatcagagt gtcgcatagt gatactaatt ctagcaaacc
      961 caaaaacacg aaggagaacc tttcaaaaag ctcttggagg caagaatggc ttgcaaattt
     1021 gaaacttatt tccgtttcgt tggttgatga gttcccttcg gagctttccg acagtgatag
     1081 acaaataatt aacgaaaaaa tgcagttact taaagatata tttgctaaca accttaaatc
     1141 agcaatttcc aataatttta gagagagtga catcattata ctgaaaggtg aaatagaaga
     1201 ttacccaatg agttctgaaa ttaagattta ctacaacgaa ttacagaaca agcctgatgc
     1261 aaaaaaagcc agattttggt catttatgaa gactcagaga tttgtttcca acatgggatt
     1321 tgatattcag aagtcctgtg aacctgtttc tatatccacg tcggtcaaac cacatgtggt
     1381 cgagcccgag catatggcag atgccaaaat tatgcctaag gatatactgc aaattacaaa
     1441 aaaaccgtta atggttaaaa atgtgaagcc ttcttctccg ccagatgtga agtctttggt
     1501 gcagctgagc acaatggaaa cgaaaaccct accagaaaag aagcaattcg acagtatttt
     1561 taattctaac aaagcaaaaa taatccctgg aaatgggaag cacgcatcag aaaacatctc
     1621 actctctttc tcaagacctg cgtcctacgg ctatttttct gttggaaaaa gggttcgggg
     1681 atcctctaga ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct
     1741 aatctccgag gaacacttga acggtgaaca aaaattaatc tcagaagaag acttgaacgg
     1801 atcctctaga gtcgactgaa tcgtagatac tgaaaaaccc cgcaagttca cttcaactgt
     1861 gcatcgtgca ccatctcaat ttctttcatt tatacatcgt tttgccttct tttatgtaac
     1921 tatactcctc taagtttcaa tcttggccat gtaacctctg atctatagaa ttttttaaat
     1981 gactagaatt aatgcccatc ttttttttgg acctaaattc ttcatgaaaa tatattacga
     2041 gggcttattc agaagctttg gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg
     2101 ttgtcggctt gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg
     2161 gtagatacgt tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta
     2221 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2281 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     2341 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccgagcaaa tgcctgcaaa
     2401 tcgctcccca tttcacccaa ttgtagatat gctaactcca gcaatgagtt gatgaatctc
     2461 ggtgtgtatt ttatgtcctc agaggacaac acctgttgta atcgttcttc cacacggatc
     2521 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2581 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2641 catatatcgc tgggccattc tcatgaagaa tatcttgaat ttattgtcat attactagtt
     2701 ggtgtggaag tccatatatc ggtgatcaat atagtggttg acatgctggc tagtcaacat
     2761 tgagcctttt gatcatgcaa atatattacg gtattttaca atcaaatatc aaacttaact
     2821 attgacttta taacttattt aggtggtaac attcttataa aaaagaaaaa aattactgca
     2881 aaacagtact agcttttaac ttgtatccta ggttatctat gctgtctcac catagagaat
     2941 attacctatt tcagaatgta tgtccatgat tcgccgggta aatacatata atacacaaat
     3001 ctggcttaat aaagtctata atatatctca taaagaagtg ctaaattggc tagtgctata
     3061 tatttttaag aaaatttctt ttgactaagt ccatatcgac tttgtaaaag ttcactttag
     3121 catacatata ttacacgagc cagaaattgt aacttttgcc taaaatcaca aattgcaaaa
     3181 tttaattgct tgcaaaaggt cacatgctta taatcaactt ttttaaaaat ttaaaatact
     3241 tttttatttt ttatttttaa acataaatga aataatttat ttattgttta tgattaccga
     3301 aacataaaac ctgctcaaga aaaagaaact gttttgtcct tggaaaaaaa gcactaccta
     3361 ggagcggcca aaatgccgag gctttcatag cttaaactct ttacagaaaa taggcattat
     3421 agatcagttc gagttttctt attcttcctt ccggttttat cgtcacagtt ttacagtaaa
     3481 taagtatcac ctcttagagt tcgatgataa gctgtcaaac atgagaatta attccacatg
     3541 ttaaaatagt gaaggagcat gttcggcaca cagtggaccg aacgtggggt aagtgcacta
     3601 gggtccggtt aaacggatct cgcattgatg aggcaacgct aattatcaac atatagattg
     3661 ttatctatct gcatgaacac gaaatcttta cttgacgact tgaggctgat ggtgtttatg
     3721 caaagaaacc actgtgttta atatgtgtca ctgtttgata ttactgtcag cgtagaagat
     3781 aatagtaaaa gcggttaata agtgtatttg agataagtgt gataaagttt ttacagcgaa
     3841 aagacgataa atacaagaaa atgattacga ggatacggag agaggtatgt acatgtgtat
     3901 ttatatacta agctgccggc ggttgtttgc aagaccgaga aaaggctagc aagaatcggg
     3961 tcattgtagc gtatgcgcct gtgaacattc tcttcaacaa gtttgattcc attgcggtga
     4021 aatggtaaaa gtcaaccccc tgcgatgtat attttcctgt acaatcaatc aaaaagccaa
     4081 atgatttagc attatcttta catcttgtta ttttacagat tttatgttta gatcttttat
     4141 gcttgctttt caaaaggctt gcaggcaagt gcacaaacaa tacttaaata aatactactc
     4201 agtaataacc tatttcttag catttttgac gaaatttgct attttgttag agtcttttac
     4261 accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta atctaagcgc
     4321 atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc tctcggggct
     4381 ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac ctgtcccacc
     4441 tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg cactgagtag
     4501 tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga ggaactcttg
     4561 gtattcttgc cacgactcat ctccatgcag ttggacgata tcaatgccgt aatcattgac
     4621 cagagccaaa acatcctcct taggttgatt acgaaacacg ccaaccaagt atttcggagt
     4681 gcctgaacta tttttatatg cttttacaag acttgaaatt ttccttgcaa taaccgggtc
     4741 aattgttctc tttctattgg gcacacatat aatacccagc aagtcagcat cggaatctag
     4801 tgcacattct gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg gaccagaact
     4861 acctgtgaaa ttaataacag acatactcca agctgccttt gtgtgcttaa tcacgtatac
     4921 tcacgtgctc aatagtcacc aatgccctcc ctcttggccc tctccttttc ttttttcgac
     4981 cgaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5041 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6361 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6841 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6901 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6961 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7021 ctccccgcgc gttggccgat tcattaatgc aggatccggg atcgaagaaa tgatggtaaa
     7081 tgaaatagga aatcaaggag catgaaggca aaagacaaat ataagggtcg aacgaaaaat
     7141 aaagtgaaaa gtgttgatat gatgtatttg gctttgcggc gccgaaaaaa cgagtttacg
     7201 caattgcaca atcatgctga ctctgtggcg gacccgcgct cttgccggcc cggcgataac
     7261 gctgggcgtg aggctgtgcc cggcggagtt ttttgcgcct gcattttcca aggtttaccc
     7321 tgcgctaagg ggcgagattg gagaagcaat aagaatgccg gttggggttg cgatgatgac
     7381 gaccacgaca actggtgtca ttatttaagt tgccgaaaga acctgagtgc atttgcaaca
     7441 tgagtatact agaagaatga gccaagactt gcgagacgcg agtttgccgg tggtgcgaac
     7501 aatagagcga ccatgacctt gaaggtgaga cgcgcataac cgctagagta ctttgaagag
     7561 gaaacagcaa tagggttgct accagtataa atagacaggt acatacaaca ctggaaatgg
     7621 ttgtctgttt gagtacgctt tcaattcatt tgggtgtgca ctttattatg ttacaatatg
     7681 gaagggaact ttacacttct cctatgcaca tatattaatt aaagtccaat gctagtagag
     7741 aaggggggta acacccctcc gcgctctttt ccgatttttt tctaaaccgt ggaatatttc
     7801 ggatatcctt ttgttgtttc cgggtgtaca atatggactt cctcttttct ggcaaccaaa
     7861 cccatacatc gggattccta taataccttc gttggtctcc ctaacatgta ggtggcggag
     7921 gggagatata caatagaaca gataccagac aagacataat gggctaaaca agactacacc
     7981 aattacactg cctcattgat g
//
