LOCUS       sequence-381351-       10267 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transcription factor tagged with Sir4 used for Calling Cards 
            experiments for binding measurement.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10267)
  TITLE     A Novel Mode of Half Site Binding in Budding Yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: ADH1pro_LEU3_SIR4
FEATURES             Location/Qualifiers
     source          1..10267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3962..3991
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3998..4027
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4034..4063
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4458..4645
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4513..4537)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    5475..6637
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      5800..6637
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(6476..7150)
                     /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(7151..7252)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7272..7290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7358..7462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7463..7531,7532..8323)
                     /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(7681..7700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8494..9082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8983..9002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9236..9253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(10071..10267,1..508)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
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 ccttcattca
      301 cgcacactac tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga
      361 aataaaaaaa agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt
      421 ttgtttcctc gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt
      481 tcaagctata ccaagcatac aatcaactcc aagcttgaag caagcctcct gaaagatgga
      541 aggaagatca gattttgtgg cgacttcaca gtccggaagt gaaatgagcc atagtgaaac
      601 taggaataga actgggatga atgctaggaa aaggaaattc gcctgtgtgg aatgtcgtca
      661 gcagaagtcg aaatgtgatg ctcacgaaag agcaccggag ccatgcacta agtgtgctaa
      721 aaagaatgtc ccatgcattt taaaacgaga tttcagaaga acttataaaa gagcaaggaa
      781 cgaagccatt gaaaaaagat tcaaggaact caccagaact ttgacaaatt taacttcgga
      841 tgaaattttg aagaaaattg aagaggaaca ggaaattgtt ttggataaca gtaatttcac
      901 aaaagaaaaa gtaaaacaac tcaggaagag tgcttttgag acgacagaaa tagaaccgag
      961 atcatacaaa acacttcgag gagaacctat ttcttacagt accaacagaa gacatacgga
     1021 ttcttctcct ttaacgctct taagctcgtc gacaaacttc gaccctgttc actcaacaaa
     1081 cgttatgaca gatgatcaac ttaagtgctt gccaaaaagc ctgggcgacg tatatttgtc
     1141 aagcagcgat attgctgagc tgtttcaaga atttgcgaca aaatatcatc aatttttacc
     1201 cgtcgttgac ctttcgaaag gagcagagcg aatttatcac ttatctcctt gcttattctg
     1261 ggtcatcctg ctcattggtt taaggcggaa atttggggct acagacttaa tgactcgatt
     1321 atcagtgcta gtaaagtcag ttttatcaga aatcacaata tctccaataa ttcgatatac
     1381 tccatcagat aaggacgaac ccgttctaaa tgtagcatct gtatattccg tgcaagcatt
     1441 tcttttatac acgttctggc ctcccttaac ctcttcatta agcgccgaca cttcgtggaa
     1501 taccatagga acagcgatgt tccaagcgct tcgggtagga ctaaattgtg caggtttttc
     1561 aaaagagtat gcttcggcaa attcagaatt agttaacgag caaatacgaa cttggatttg
     1621 ctgcaatgtt gtatctcaaa cagttgcatc atcatttggt ttcccagctt atgtttcatt
     1681 tgattattta gtaatcagct ctattagagt accaaattca aaaagccaag tagatatacc
     1741 caatgaacta agacaaatgg ctcaaattgc tagatttgag aaccaaatcg taaacacaat
     1801 gaactccacc ccggcgagtg ttactgggat ggtaagtcag gaagagaagc agcccttgtt
     1861 acacgttctt aatcaacaac taagtcaatt ggagattagt cttgaagaaa ataacctaga
     1921 tgatatccga aaatttttat tactagtggc caaggttcac ttattaacct attacttcac
     1981 tgatgttacc tcccaaagtg ctggaaaatc aaatggtaat atttatgagg gatcatattc
     2041 cattatggaa ctcgatacaa gttttgaaac gaaacgtgga ttggtgaaag tttataatgc
     2101 cgccgtaaac tttcttatac atgccaatag tatgtgggaa catgatccta ccattattaa
     2161 gtactttcct ggtttgtttg tcttgaatat atggcaatct gcctgtatta ttagtaaact
     2221 catacattca tcacttcatt caatgctaga tgttaactca ggcaaaaaag cttataacaa
     2281 cgcaatttca ttgacgttta atgcctcagt tttaaaatat gatatggcgt acagatcatc
     2341 cggaataatg cgaagcatat ggagtttatt tgctaatatg tatgatgcct ggaaaaacga
     2401 ccaaaaggaa ggtggaggta gactaaataa tgatttcaat ttaggcatca ccataaaatc
     2461 taggatgtca gtaaacgttt tttttgactg cttatatatt ctaaaagaga aatgtggtat
     2521 ggccaaattg gagagagaga ccaaggtttc tacagcttac aatgttgatg aagaggaaga
     2581 ggaagatgaa gatgaggagg gagaagaaga agaagaagaa gaagaactaa gtagtaaagt
     2641 tccagaaaat atggatagcc agcaactaag gacaaggaaa ttcaccaatg taaggcatcc
     2701 agaaaagaaa gcaagaaaaa taattgaaac aattccgcta gacccaaatc caataaatgc
     2761 aggctctacc agcagtggaa gctcattaac gaccccaaat agtcaagtag cgaacactat
     2821 atcatataga ggaatcctca ataaaatgtc acctagggaa caactgaatc atgcaaattt
     2881 agattccagt gtttctacag acatcaagga cactgaagct gtcaatgaac ctctgccaat
     2941 agggaggaat gctgaacatc cggcaaatca accgcctctt tcaataacac aaatgcaaga
     3001 aaacacacta cctgcgacac aagccaactc ttctctatta gaaacgtatc ccattgttca
     3061 atcaaacccc gttacaacta caatcaaaga atcacccaat tccatcatgg caggttggga
     3121 taactgggaa tctgatatgg tttggaggga tgttgatatt ttaatgaatg aatttgcgtt
     3181 caatcccaag gttagagtgt cgcatagtga tactaattct agcaaacccg aaaacacgaa
     3241 ggagaacctt tcaaaaagct cttggaggca ggaatggctt gcaaatttga aacttatttc
     3301 cgtttcgttg gttgatgagt tcccttcgga gctttccgac agtgatagac aagtaattaa
     3361 cgaaaaaatg cagttactta aagatatatt tgctaacaac cttaaatcag caatttccaa
     3421 taattttaga gagagtgaca tcattatact gaaaggtgaa atagaagatt acccaatgag
     3481 ttctgaaatt aagatttact acaacgaatt acagaacaag cctgatgcaa aaaaagccag
     3541 attttggtca tttatgaaga ctcagagatt tgtttccaac atgggatttg atattcggaa
     3601 gtcctgtgaa cctgtttcta tatccacgtc ggtcaaacca catgtggtcg agcccgagca
     3661 tatggcagat gccaaaatta tgcctaagga tgtactgcaa attacaaaaa aaccgttaat
     3721 ggttaaaaat gtgaagcctt cttctccgcc agatgtgaag tctttggtgc agctgagcac
     3781 aatggaaacg aaaaccctac cagaaaagaa gcaattcgac agtattttta attctaacaa
     3841 agcaaaaata atccctggaa atgggaagca cgcatcagaa aacatctcac tctctttctc
     3901 aagacctgcg tcctacggct atttttctgt tggaaaaagg gttcggggat cctctagagg
     3961 tgaacaaaag ttgatttctg aagaagattt gaacggtgaa caaaagctaa tctccgagga
     4021 agacttgaac ggtgaacaaa aattaatctc agaagaagac ttgaacggat cctctagagt
     4081 cgactgaatc gtagatactg aaaaaccccg caagttcact tcaactgtgc atcgtgcacc
     4141 atctcaattt ctttcattta tacatcgttt tgccttcttt tatgtaacta tactcctcta
     4201 agtttcaatc ttggccatgt aacctctgat ctatagaatt ttttaaatga ctagaattaa
     4261 tgcccatctt ttttttggac ctaaattctt catgaaaata tattacgagg gcttattcag
     4321 aagctttgga cttcttcgcc agaggtttgg tcaagtctcc aatcaaggtt gtcggcttgt
     4381 ctaccttgcc agaaatttac gaaaagatgg aaaagggtca aatcgttggt agatacgttg
     4441 ttgacacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta
     4501 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct
     4561 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg
     4621 ctcttattga ccacacctct accggcatgc cgagcaaatg cctgcaaatc gctccccatt
     4681 tcacccaatt gtagatatgc taactccagc aatgagttga tgaatctcgg tgtgtatttt
     4741 atgtcctcag aggacaacac ctgttgtaat cgttcttcca cacggatcct ggcgtaatag
     4801 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4861 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taatcgctgg
     4921 gccattctca tgaagaatat cttgaattta ttgtcatatt actagttggt gtggaagtcc
     4981 taatatcggt gatcaatata gtggttgaca tgctggctag tcaacattga gccttttgat
     5041 catgcaaata tattacggta ttttacaatc aaatatcaaa cttaactatt gactttataa
     5101 cttatttagg tggtaacatt cttataaaaa agaaaaaaat tactgcaaaa cagtactagc
     5161 ttttaacttg tatcctaggt tatctatgct gtctcaccat agagaatatt acctatttca
     5221 gaatgtatgt ccatgattcg ccgggtaaat acatataata cacaaatctg gcttaataaa
     5281 gtctataata tatctcataa agaagtgcta aattggctag tgctatatat ttttaagaaa
     5341 atttcttttg actaagtcca tatcgacttt gtaaaagttc acattagcat acatatatta
     5401 cacgagccag aaatagtaac ttttgcctaa atcacaaatt gcaaaattta attgcttgca
     5461 aaaggtcaca tgcttataat caactttttt aaaaatttaa aatacttttt tattttttat
     5521 ttttaaacat aaatgaaata atttatttat tgtttatgat taccgaaaca taaaacctgc
     5581 tcaagaaaaa gaaactgttt tgtccttgga aaaaaagcac tacctaggag cggccaaaat
     5641 gccgaggctt tcatagctta aactctttac agaaaatagg cattatagat cagttcgagt
     5701 tttcttattc ttccttccgg ttttatcgtc acagttttac agtaaataag tatcacctct
     5761 tagagttcga tgataagctg tcaaacatga gaattaattc cacatgttaa aatagtgaag
     5821 gagcatgttc ggcacacagt ggaccgaacg tggggtaagt gcactagggt ccggttaaac
     5881 ggatctcgca ttgatgaggc aacgctaatt atcaacatat agattgttat ctatctgcat
     5941 gaacacgaaa tctttacttg acgacttgag gctgatggtg tttatgcaaa gaaaccactg
     6001 tgtttaatat gtgtcactgt ttgatattac tgtcagcgta gaagataata gtaaaagcgg
     6061 ttaataagtg tatttgagat aagtgtgata aagtttttac agcgaaaaga cgataaatac
     6121 aagaaaatga ttacgaggat acggagagag gtatgtacat gtgtatttat atactaagct
     6181 gccggcggtt gtttgcaaga ccgagaaaag gctagcaaga atcgggtcat tgtagcgtat
     6241 gcgcctgtga acattctctt caacaagttt gattccattg cggtgaaatg gtaaaagtca
     6301 accccctgcg atgtatattt tcctgtacaa tcaatcaaaa agccaaatga tttagcatta
     6361 tctttacatc ttgttatttt acagatttta tgtttagatc ttttatgctt gcttttcaaa
     6421 aggcttgcag gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt
     6481 tcttagcatt tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca
     6541 cacctccgct tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt
     6601 ctggcgtcag tccaccagct aacataaaat gtaagctctc ggggctctct tgccttccaa
     6661 cccagtcaga aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa
     6721 acaagggaat aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt
     6781 ttggaaatac gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg
     6841 actcatctcc atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat
     6901 cctccttagg ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt
     6961 tatatgcttt tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc
     7021 tattgggcac acatataata cccagcaagt cagcatcgga atctagtgca cattctgcgg
     7081 cctctgtgct ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa
     7141 taacagacat actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata
     7201 gtcaccaatg ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattcttg
     7261 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7321 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7381 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7441 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7561 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7621 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7681 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7741 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7861 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7981 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8041 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8281 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8641 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     9181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     9241 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     9301 gccgattcat taatgcagga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
     9361 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
     9421 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
     9481 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
     9541 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
     9601 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
     9661 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
     9721 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
     9781 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
     9841 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
     9901 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
     9961 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
    10021 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
    10081 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
    10141 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
    10201 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
    10261 attgatg
//
