LOCUS       sequence-274988-       10595 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 10595)
  TITLE     Mitra lab plasmids
REFERENCE   2  (bases 1 to 10595)
  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_GCR2_SIR4
FEATURES             Location/Qualifiers
     source          1..10595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2398..2427
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2434..2463
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2470..2499
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      2894..3081
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2949..2973)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    3911..4203
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     CDS             complement(5328..6422)
                     /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(6423..6830)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(7092..7110)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7178..7282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7283..7351,7352..8143)
                     /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(7501..7520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8314..8902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8803..8822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9056..9073
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        9891..10595
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 ccaagcttga agcaagcctc ctgaaagatg catcaccaaa ctaagttaga tgtattcata
       61 atcagagctt ataatttact gtctaacgag tctgtcatta gtggtgcttc cttgcagagt
      121 gttacaaact cgccacagac gacaacgaac acgccctcag gtatggttaa cggggcggtt
      181 ggaacaggga tagctaatcc aacagggttg atggggtctg atagcacacc taacatcgat
      241 gagattataa ctagcactgg tagtaatgct ctgacgaaaa ccaactcaga tagcgctaat
      301 ggtacgccga atggtaattc aagttctacc tcagccatta gcaatgcaag caatcctgcc
      361 actactggta ataatgcgag ctctagtgcc acctcaaatg gaatatatac gcaagcgcaa
      421 tattctcaac ttttcgccaa aatatcaaaa ctatataacg ctacactatc atctgggtca
      481 attgacgata gatcaacatc accaaaatcg gcaatcgaac tatatcaaag atttcaacag
      541 atgattaagg aactagagct gagttttgac gcaagtcctt acgcaaaata cttccgccgg
      601 ttggatggaa ggctttggca aataaagaca gactcagaat tagaaaacga tgaattgtgg
      661 cgattagtct caatgagcat atttacagta ttcgatcctc agaccggcca aattctaact
      721 caaggacgca ggaagggaaa ctccttaaat acatcaacta aaggctcccc atcagattta
      781 cagggaataa acaacgggaa caataatggg aacaatggta atattggaaa tgggagtaat
      841 attaagaact atggaaataa aaacatgcca aacaaccgaa cgaaaaaaag aggcaccagg
      901 gtggctaaaa atgctaaaaa tgggaaaaac aataaaaata gtaataaaga gagaaacggc
      961 attacagata cgagtgcatt cagtaataca acaataagca acccaggtac caatatgctt
     1021 tttgatccat cattgtctca acagttacaa aaacgactgc aaacgctatc acaagatgtc
     1081 aattctcgtt cgttgacagg atattataca cagccaacca gtcctggctc aggaggattt
     1141 gaatttggtt tgagtcatgc agatctgaac ccaaatgctt ccagtaatac catgggctat
     1201 aatacaatgt ccaataatgg atcccattcg tggaaacgaa ggtcactggg atcgttagat
     1261 gttaatacgc tggatgacga agcggtggaa gaacttttgc aactgacaaa tacgagtaag
     1321 aggcagaggc cgatgacaac tgcagcagag ggtgcgttaa taaatgatgg tccggacact
     1381 aatttaaacg cgaataacac ccaaatgaaa gttgatttaa atccttcaaa cagcatggga
     1441 cctatagata cagaagccgt gatacgccca ttgaaagaag cttatgacgc aatcatttct
     1501 gaaaaaggcc aaagaattgt gcaattagaa agagaattgg aattacagcg ccaagagacg
     1561 cagtggttaa ggaaaatgtt aattgaagac atgggttgtg ttagaagtat gttaagggat
     1621 ttacaaagaa gagtgtcgca tagtgatact aattctagca aacccaaaaa cacgaaggag
     1681 aacctttcaa aaagctcttg gaggcaagaa tggcttgcaa atttgaaact tatttccgtt
     1741 tcgttggttg atgagttccc ttcggagctt tccgacagtg atagacaaat aattaacgaa
     1801 aaaatgcagt tacttaaaga tatatttgct aacaacctta aatcagcaat ttccaataat
     1861 tttagagaga gtgacatcat tatactgaaa ggtgaaatag aagattaccc aatgagttct
     1921 gaaattaaga tttactacaa cgaattacag aacaagcctg atgcaaaaaa agccagattt
     1981 tggtcattta tgaagactca gagatttgtt tccaacatgg gatttgatat tcagaagtcc
     2041 tgtgaacctg tttctatatc cacgtcggtc aaaccacatg tggtcgagcc cgagcatatg
     2101 gcagatgcca aaattatgcc taaggatata ctgcaaatta caaaaaaacc gttaatggtt
     2161 aaaaatgtga agccttcttc tccgccagat gtgaagtctt tggtgcagct gagcacaatg
     2221 gaaacgaaaa ccctaccaga aaagaagcaa ttcgacagta tttttaattc taacaaagca
     2281 aaaataatcc ctggaaatgg gaagcacgca tcagaaaaca tctcactctc tttctcaaga
     2341 cctgcgtcct acggctattt ttctgttgga aaaagggttc ggggatcctc tagaggtgaa
     2401 caaaagttga tttctgaaga agatttgaac ggtgaacaaa agctaatctc cgaggaagac
     2461 ttgaacggtg aacaaaaatt aatctcagaa gaagacttga acggatcctc tagagtcgac
     2521 tgaatcgtag atactgaaaa accccgcaag ttcacttcaa ctgtgcatcg tgcaccatct
     2581 caatttcttt catttataca tcgttttgcc ttcttttatg taactatact cctctaagtt
     2641 tcaatcttgg ccatgtaacc tctgatctat agaatttttt aaatgactag aattaatgcc
     2701 catctttttt ttggacctaa attcttcatg aaaatatatt acgagggctt attcagaagc
     2761 tttggacttc ttcgccagag gtttggtcaa gtctccaatc aaggttgtcg gcttgtctac
     2821 cttgccagaa atttacgaaa agatggaaaa gggtcaaatc gttggtagat acgttgttga
     2881 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     2941 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     3001 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     3061 tattgaccac acctctaccg gcatgccgag caaatgcctg caaatcgctc cccatttcac
     3121 ccaattgtag atatgctaac tccagcaatg agttgatgaa tctcggtgtg tattttatgt
     3181 cctcagagga caacacctgt tgtaatcgtt cttccacacg gatcctggcg taatagcgaa
     3241 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     3301 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataat cgctgggcca
     3361 ttctcatgaa gaatatcttg aatttattgt catattacta gttggtgtgg aagtcctaat
     3421 atcggtgatc aatatagtgg ttgacatgct ggctagtcaa cattgagcct tttgatcatg
     3481 caaatatatt acggtatttt acaatcaaat atcaaactta actattgact ttataactta
     3541 tttaggtggt aacattctta taaaaaagaa aaaaattact gcaaaacagt actagctttt
     3601 aacttgtatc ctaggttatc tatgctgtct caccatagag aatattacct atttcagaat
     3661 gtatgtccat gattcgccgg gtaaatacat ataatacaca aatctggctt aataaagtct
     3721 ataatatatc tcataaagaa gtgctaaatt ggctagtgct atatattttt aagaaaattt
     3781 cttttgacta agtccatatc gactttgtaa aagttcacat tagcatacat atattacacg
     3841 agccagaaat agtaactttt gcctaaatca caaattgcaa aatttaattg cttgcaaaag
     3901 gtcacatgct tataatcaac ttttttaaaa atttaaaata cttttttatt ttttattttt
     3961 aaacataaat gaaataattt atttattgtt tatgattacc gaaacataaa acctgctcaa
     4021 gaaaaagaaa ctgttttgtc cttggaaaaa aagcactacc taggagcggc caaaatgccg
     4081 aggctttcat agcttaaact ctttacagaa aataggcatt atagatcagt tcgagttttc
     4141 ttattcttcc ttccggtttt atcgtcacag ttttacagta aataagtatc acctcttaga
     4201 gttcgatgat aagctgtcaa acatgagaat taattccaca tgttaaaata gtgaaggagc
     4261 atgttcggca cacagtggac cgaacgtggg gtaagtgcac tagggtccgg ttaaacggat
     4321 ctcgcattga tgaggcaacg ctaattatca acatatagat tgttatctat ctgcatgaac
     4381 acgaaatctt tacttgacga cttgaggctg atggtgttta tgcaaagaaa ccactgtgtt
     4441 taatatgtgt cactgtttga tattactgtc agcgtagaag ataatagtaa aagcggttaa
     4501 taagtgtatt tgagataagt gtgataaagt ttttacagcg aaaagacgat aaatacaaga
     4561 aaatgattac gaggatacgg agagaggtat gtacatgtgt atttatatac taagctgccg
     4621 gcggttgttt gcaagaccga gaaaaggcta gcaagaatcg ggtcattgta gcgtatgcgc
     4681 ctgtgaacat tctcttcaac aagtttgatt ccattgcggt gaaatggtaa aagtcaaccc
     4741 cctgcgatgt atattttcct gtacaatcaa tcaaaaagcc aaatgattta gcattatctt
     4801 tacatcttgt tattttacag attttatgtt tagatctttt atgcttgctt ttcaaaaggc
     4861 ttgcaaggcc gtttctgaca gagtaaaatt cttgagggaa ctttcaccat tatgggaaat
     4921 ggttcaagaa ggtattgact taaactccat caaatggtca ggtcattgag tgttttttat
     4981 ttgttgtatt tttttttttt tagagaaaat cctccaatat ataaattagg aatcatagtt
     5041 tcatgatttt ctgttacacc taactttttg tgtggtgccc tcctccttgt caatattaat
     5101 gttaaagtgc aattcttttt ccttatcacg ttgagccatt agtatcaatt tgcttacctg
     5161 tattccttta catcctcctt tttctccttc ttgataaatg tatgtagatt gcgtatatag
     5221 tttcgtctac cctatgaaca tattccattt tgtaatttcg tgtcgtttct attatgaatt
     5281 tcatttataa agtttatgta caaatatcat aaaaaaagag aatcttttta agcaaggatt
     5341 ttcttaactt cttcggcgac agcatcaccg acttcggtgg tactgttgga accacctaaa
     5401 tcaccagttc tgatacctgc atccaaaacc tttttaactg catcttcaat ggccttacct
     5461 tcttcaggca agttcaatga caatttcaac atcattgcag cagacaagat agtggcgata
     5521 gggttgacct tattctttgg caaatctgga gcagaaccgt ggcatggttc gtacaaacca
     5581 aatgcggtgt tcttgtctgg caaagaggcc aaggacgcag atggcaacaa acccaaggaa
     5641 cctgggataa cggaggcttc atcggagatg atatcaccaa acatgttgct ggtgattata
     5701 ataccattta ggtgggttgg gttcttaact aggatcatgg cggcagaatc aatcaattga
     5761 tgttgaacct tcaatgtagg gaattcgttc ttgatggttt cctccacagt ttttctccat
     5821 aatcttgaag aggccaaaac attagcttta tccaaggacc aaataggcaa tggtggctca
     5881 tgttgtaggg ccatgaaagc ggccattctt gtgattcttt gcacttctgg aacggtgtat
     5941 tgttcactat cccaagcgac accatcacca tcgtcttcct ttctcttacc aaagtaaata
     6001 cctcccacta attctctgac aacaacgaag tcagtacctt tagcaaattg tggcttgatt
     6061 ggagataagt ctaaaagaga gtcggatgca aagttacatg gtcttaagtt ggcgtacaat
     6121 tgaagttctt tacggatttt tagtaaacct tgttcaggtc taacactacc ggtaccccat
     6181 ttaggaccac ccacagcacc taacaaaacg gcatcagcct tcttggaggc ttccagcgcc
     6241 tcatctggaa gtggaacacc tgtagcatcg atagcagcac caccaattaa atgattttcg
     6301 aaatcgaact tgacattgga acgaacatca gaaatagctt taagaacctt aatggcttcg
     6361 gctgtgattt cttgaccaac gtggtcacct ggcaaaacga cgatcttctt aggggcagac
     6421 attagaatgg tatatccttg aaatatatat atatatattg ctgaaatgta aaaggtaaga
     6481 aaagttagaa agtaagacga ttgctaacca cctattggaa aaaacaatag gtccttaaat
     6541 aatattgtca acttcaagta ttgtgatgca agcatttagt catgaacgct tctctattct
     6601 atatgaaaag ccggttccgg cgctctcacc tttccttttt ctcccaattt ttcagttgaa
     6661 aaaggtatat gcgtcaggcg acctctgaaa ttaacaaaaa atttccagtc atcgaatttg
     6721 attctgtgcg atagcgcccc tgtgtgttct cgttatgttg aggaaaaaaa taatggttgc
     6781 taagagattc gaactcttgc atcttacgat acctgagtat tcccacagtt aactgcggtc
     6841 aagatatttc ttgaatcagg cgccttagac cgctcggcca aacaaccaat tacttgttga
     6901 gaaatagagt ataattatcc tataaatata acgtttttga acacacatga acaaggaagt
     6961 acaggacaat tgattttgaa gagaatgtgg attttgatgt aattgttggg attccatttt
     7021 taataaggca ataatattag gtatgtagat atactagaag ttctcctcga ggaattcttg
     7081 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7141 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7201 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7261 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7321 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7381 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7441 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7501 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7561 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7621 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7681 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7741 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7801 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7861 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7921 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7981 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8041 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8101 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8161 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8221 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8461 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     9001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     9061 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     9121 gccgattcat taatgcagga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
     9181 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
     9241 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
     9301 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
     9361 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
     9421 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
     9481 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
     9541 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
     9601 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
     9661 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
     9721 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
     9781 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
     9841 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
     9901 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
     9961 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
    10021 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
    10081 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
    10141 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
    10201 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
    10261 gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag
    10321 atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct
    10381 tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc cagttacttg
    10441 aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat
    10501 taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca
    10561 caatacttca agctatacca agcatacaat caact
//
