LOCUS       sequence-274925-        9866 bp    DNA     circular SYN 15-DEC-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 9866)
  TITLE     Mitra lab plasmids
REFERENCE   2  (bases 1 to 9866)
  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"
COMMENT     Sequence Label: ADH1pro_TYE7_SIR4
FEATURES             Location/Qualifiers
     source          1..9866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..705
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             2374..2403
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2410..2439
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2446..2475
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2870..3057
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2925..2949)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    3887..4179
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     CDS             complement(5304..6398)
                     /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(6399..6806)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(7068..7086)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7154..7258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7259..7327,7328..8119)
                     /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(7477..7496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8290..8878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8779..8798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9032..9049
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atccttttgt tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca
       61 tacatcggga ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga
      121 gatatacaat agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt
      181 acactgcctc attgatggtg gtacataacg aactaatact gtagccctag acttgatagc
      241 catcatcata tcgaagtttc actacccttt ttccatttgc catctattga agtaataata
      301 ggcgcatgca acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca
      361 tatccgcaat gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca
      421 gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt
      481 tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc
      541 cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac
      601 ctgcaattat taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc
      661 tttttctgca caatacttca agctatacca agcatacaat caactccaag cttgaagcaa
      721 gcctcctgaa agatgaactc tattttagac agaaatgtta gatctagcga aactacttta
      781 attaaacctg aatctgaatt tgataattgg ttgtcggatg aaaatgacgg agctagtcat
      841 atcaacgtca acaaggactc ctcgtcagtt ctttctgcat cttcttccac atggttcgaa
      901 ccattggaaa acattatctc ctctgcatcc agctcctcga taggctctcc aatcgaagac
      961 cagtttatat cttccaacaa cgaggaatct gctctttttc caacagatca gtttttcagt
     1021 aatccttcct catactcgca ttctcccgag gttagcagct cgataaaaag agaagaggat
     1081 gacaatgccc tttcgttggc agattttgaa ccggcttctt tgcaattaat gcctaacatg
     1141 ataaatactg ataataatga cgatagtacc ccacttaaga atgaaatcga gctaaacgac
     1201 tcgtttataa aaacaaatct agatgctaag gaaacgaaaa agagggctcc aagaaaaaga
     1261 ctgaccccct tccaaaagca agctcacaac aagattgaaa aacgctacag aataaacatc
     1321 aacacaaaga ttgcaagact gcagcagatt atcccatggg tagcaagtga acaaacagct
     1381 ttcgaagtag gtgattctgt aaaaaaacag gacgaagacg gcgcagaaac tgccgctact
     1441 actcctcttc catctgccgc tgctacaagc acgaagctaa ataaaagcat gatcctagaa
     1501 aaagctgttg actatattct atatctacaa aataacgaac gactatacga aatggaagtt
     1561 caaaggttga aaagtgaaat cgacactttg aaacaagacc aaaaaagagt gtcgcatagt
     1621 gatactaatt ctagcaaacc caaaaacacg aaggagaacc tttcaaaaag ctcttggagg
     1681 caagaatggc ttgcaaattt gaaacttatt tccgtttcgt tggttgatga gttcccttcg
     1741 gagctttccg acagtgatag acaaataatt aacgaaaaaa tgcagttact taaagatata
     1801 tttgctaaca accttaaatc agcaatttcc aataatttta gagagagtga catcattata
     1861 ctgaaaggtg aaatagaaga ttacccaatg agttctgaaa ttaagattta ctacaacgaa
     1921 ttacagaaca agcctgatgc aaaaaaagcc agattttggt catttatgaa gactcagaga
     1981 tttgtttcca acatgggatt tgatattcag aagtcctgtg aacctgtttc tatatccacg
     2041 tcggtcaaac cacatgtggt cgagcccgag catatggcag atgccaaaat tatgcctaag
     2101 gatatactgc aaattacaaa aaaaccgtta atggttaaaa atgtgaagcc ttcttctccg
     2161 ccagatgtga agtctttggt gcagctgagc acaatggaaa cgaaaaccct accagaaaag
     2221 aagcaattcg acagtatttt taattctaac aaagcaaaaa taatccctgg aaatgggaag
     2281 cacgcatcag aaaacatctc actctctttc tcaagacctg cgtcctacgg ctatttttct
     2341 gttggaaaaa gggttcgggg atcctctaga ggtgaacaaa agttgatttc tgaagaagat
     2401 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
     2461 tcagaagaag acttgaacgg atcctctaga gtcgactgaa tcgtagatac tgaaaaaccc
     2521 cgcaagttca cttcaactgt gcatcgtgca ccatctcaat ttctttcatt tatacatcgt
     2581 tttgccttct tttatgtaac tatactcctc taagtttcaa tcttggccat gtaacctctg
     2641 atctatagaa ttttttaaat gactagaatt aatgcccatc ttttttttgg acctaaattc
     2701 ttcatgaaaa tatattacga gggcttattc agaagctttg gacttcttcg ccagaggttt
     2761 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
     2821 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     2881 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2941 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     3001 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     3061 gccgagcaaa tgcctgcaaa tcgctcccca tttcacccaa ttgtagatat gctaactcca
     3121 gcaatgagtt gatgaatctc ggtgtgtatt ttatgtcctc agaggacaac acctgttgta
     3181 atcgttcttc cacacggatc ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3241 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3301 ctgtgcggta tttcacaccg cataatcgct gggccattct catgaagaat atcttgaatt
     3361 tattgtcata ttactagttg gtgtggaagt cctaatatcg gtgatcaata tagtggttga
     3421 catgctggct agtcaacatt gagccttttg atcatgcaaa tatattacgg tattttacaa
     3481 tcaaatatca aacttaacta ttgactttat aacttattta ggtggtaaca ttcttataaa
     3541 aaagaaaaaa attactgcaa aacagtacta gcttttaact tgtatcctag gttatctatg
     3601 ctgtctcacc atagagaata ttacctattt cagaatgtat gtccatgatt cgccgggtaa
     3661 atacatataa tacacaaatc tggcttaata aagtctataa tatatctcat aaagaagtgc
     3721 taaattggct agtgctatat atttttaaga aaatttcttt tgactaagtc catatcgact
     3781 ttgtaaaagt tcacattagc atacatatat tacacgagcc agaaatagta acttttgcct
     3841 aaatcacaaa ttgcaaaatt taattgcttg caaaaggtca catgcttata atcaactttt
     3901 ttaaaaattt aaaatacttt tttatttttt atttttaaac ataaatgaaa taatttattt
     3961 attgtttatg attaccgaaa cataaaacct gctcaagaaa aagaaactgt tttgtccttg
     4021 gaaaaaaagc actacctagg agcggccaaa atgccgaggc tttcatagct taaactcttt
     4081 acagaaaata ggcattatag atcagttcga gttttcttat tcttccttcc ggttttatcg
     4141 tcacagtttt acagtaaata agtatcacct cttagagttc gatgataagc tgtcaaacat
     4201 gagaattaat tccacatgtt aaaatagtga aggagcatgt tcggcacaca gtggaccgaa
     4261 cgtggggtaa gtgcactagg gtccggttaa acggatctcg cattgatgag gcaacgctaa
     4321 ttatcaacat atagattgtt atctatctgc atgaacacga aatctttact tgacgacttg
     4381 aggctgatgg tgtttatgca aagaaaccac tgtgtttaat atgtgtcact gtttgatatt
     4441 actgtcagcg tagaagataa tagtaaaagc ggttaataag tgtatttgag ataagtgtga
     4501 taaagttttt acagcgaaaa gacgataaat acaagaaaat gattacgagg atacggagag
     4561 aggtatgtac atgtgtattt atatactaag ctgccggcgg ttgtttgcaa gaccgagaaa
     4621 aggctagcaa gaatcgggtc attgtagcgt atgcgcctgt gaacattctc ttcaacaagt
     4681 ttgattccat tgcggtgaaa tggtaaaagt caaccccctg cgatgtatat tttcctgtac
     4741 aatcaatcaa aaagccaaat gatttagcat tatctttaca tcttgttatt ttacagattt
     4801 tatgtttaga tcttttatgc ttgcttttca aaaggcttgc aaggccgttt ctgacagagt
     4861 aaaattcttg agggaacttt caccattatg ggaaatggtt caagaaggta ttgacttaaa
     4921 ctccatcaaa tggtcaggtc attgagtgtt ttttatttgt tgtatttttt tttttttaga
     4981 gaaaatcctc caatatataa attaggaatc atagtttcat gattttctgt tacacctaac
     5041 tttttgtgtg gtgccctcct ccttgtcaat attaatgtta aagtgcaatt ctttttcctt
     5101 atcacgttga gccattagta tcaatttgct tacctgtatt cctttacatc ctcctttttc
     5161 tccttcttga taaatgtatg tagattgcgt atatagtttc gtctacccta tgaacatatt
     5221 ccattttgta atttcgtgtc gtttctatta tgaatttcat ttataaagtt tatgtacaaa
     5281 tatcataaaa aaagagaatc tttttaagca aggattttct taacttcttc ggcgacagca
     5341 tcaccgactt cggtggtact gttggaacca cctaaatcac cagttctgat acctgcatcc
     5401 aaaacctttt taactgcatc ttcaatggcc ttaccttctt caggcaagtt caatgacaat
     5461 ttcaacatca ttgcagcaga caagatagtg gcgatagggt tgaccttatt ctttggcaaa
     5521 tctggagcag aaccgtggca tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa
     5581 gaggccaagg acgcagatgg caacaaaccc aaggaacctg ggataacgga ggcttcatcg
     5641 gagatgatat caccaaacat gttgctggtg attataatac catttaggtg ggttgggttc
     5701 ttaactagga tcatggcggc agaatcaatc aattgatgtt gaaccttcaa tgtagggaat
     5761 tcgttcttga tggtttcctc cacagttttt ctccataatc ttgaagaggc caaaacatta
     5821 gctttatcca aggaccaaat aggcaatggt ggctcatgtt gtagggccat gaaagcggcc
     5881 attcttgtga ttctttgcac ttctggaacg gtgtattgtt cactatccca agcgacacca
     5941 tcaccatcgt cttcctttct cttaccaaag taaatacctc ccactaattc tctgacaaca
     6001 acgaagtcag tacctttagc aaattgtggc ttgattggag ataagtctaa aagagagtcg
     6061 gatgcaaagt tacatggtct taagttggcg tacaattgaa gttctttacg gatttttagt
     6121 aaaccttgtt caggtctaac actaccggta ccccatttag gaccacccac agcacctaac
     6181 aaaacggcat cagccttctt ggaggcttcc agcgcctcat ctggaagtgg aacacctgta
     6241 gcatcgatag cagcaccacc aattaaatga ttttcgaaat cgaacttgac attggaacga
     6301 acatcagaaa tagctttaag aaccttaatg gcttcggctg tgatttcttg accaacgtgg
     6361 tcacctggca aaacgacgat cttcttaggg gcagacatta gaatggtata tccttgaaat
     6421 atatatatat atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc
     6481 taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt
     6541 gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcgct
     6601 ctcacctttc ctttttctcc caatttttca gttgaaaaag gtatatgcgt caggcgacct
     6661 ctgaaattaa caaaaaattt ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg
     6721 tgttctcgtt atgttgagga aaaaaataat ggttgctaag agattcgaac tcttgcatct
     6781 tacgatacct gagtattccc acagttaact gcggtcaaga tatttcttga atcaggcgcc
     6841 ttagaccgct cggccaaaca accaattact tgttgagaaa tagagtataa ttatcctata
     6901 aatataacgt ttttgaacac acatgaacaa ggaagtacag gacaattgat tttgaagaga
     6961 atgtggattt tgatgtaatt gttgggattc catttttaat aaggcaataa tattaggtat
     7021 gtagatatac tagaagttct cctcgaggaa ttcttgaaga cgaaagggcc tcgtgatacg
     7081 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7201 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7261 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7321 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7381 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7441 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7501 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7561 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7621 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7681 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7741 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7801 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     7861 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     7921 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7981 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8041 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8101 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8401 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8941 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9001 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     9061 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatccg
     9121 ggatcgaaga aatgatggta aatgaaatag gaaatcaagg agcatgaagg caaaagacaa
     9181 atataagggt cgaacgaaaa ataaagtgaa aagtgttgat atgatgtatt tggctttgcg
     9241 gcgccgaaaa aacgagttta cgcaattgca caatcatgct gactctgtgg cggacccgcg
     9301 ctcttgccgg cccggcgata acgctgggcg tgaggctgtg cccggcggag ttttttgcgc
     9361 ctgcattttc caaggtttac cctgcgctaa ggggcgagat tggagaagca ataagaatgc
     9421 cggttggggt tgcgatgatg acgaccacga caactggtgt cattatttaa gttgccgaaa
     9481 gaacctgagt gcatttgcaa catgagtata ctagaagaat gagccaagac ttgcgagacg
     9541 cgagtttgcc ggtggtgcga acaatagagc gaccatgacc ttgaaggtga gacgcgcata
     9601 accgctagag tactttgaag aggaaacagc aatagggttg ctaccagtat aaatagacag
     9661 gtacatacaa cactggaaat ggttgtctgt ttgagtacgc tttcaattca tttgggtgtg
     9721 cactttatta tgttacaata tggaagggaa ctttacactt ctcctatgca catatattaa
     9781 ttaaagtcca atgctagtag agaagggggg taacacccct ccgcgctctt ttccgatttt
     9841 tttctaaacc gtggaatatt tcggat
//
