LOCUS       sequence-275117-       10046 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 10046)
  TITLE     Mitra lab plasmids
REFERENCE   2  (bases 1 to 10046)
  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_CBF1_SIR4
FEATURES             Location/Qualifiers
     source          1..10046
                     /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             2554..2583
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2590..2619
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2626..2655
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3050..3237
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3105..3129)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    4067..4359
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     CDS             complement(5484..6578)
                     /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(6579..6986)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(7248..7266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7334..7438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7439..7507,7508..8299)
                     /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(7657..7676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8470..9058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8959..8978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9212..9229
                     /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 tctggcaaat aataataagc tttctactga ggatgaggaa
      781 atccattcgg caagaaaaag aggctataac gaggagcaga actacagcga ggccaggaaa
      841 aagcaaagag atcaaggctt gttgtcacaa gagagcaatg acggaaatat tgactctgct
      901 ctgttgagcg aaggggctac gctaaaaggg actcaaagtc agtatgaaag cggactgacg
      961 tctaacaaag atgagaaagg aagtgatgac gaagatgcgt cagtggctga ggctgctgtt
     1021 gccgctaccg tcaattatac agacttaatc cagggccagg aagatagttc tgatgcccac
     1081 acatctaatc aaacgaacgc gaatggtgag cacaaggatt cccttaatgg ggaaagagcc
     1141 ataaccccat caaatgaagg agtcaagcca aacacctccc tggagggaat gacgtcttca
     1201 ccaatggaat ctacacagca atctaaaaac gacatgctca taccattggc agaacacgat
     1261 cgtgggccag aacaccaaca ggatgatgaa gacaatgatg acgctgacat tgatctcaaa
     1321 aaggatataa gcatgcagcc gggtcgtcgt ggaagaaaac ctactacttt ggccacaaca
     1381 gacgagtgga aaaagcaaag aaaagactcc cataaagaag tcgaaaggcg tcgccgcgaa
     1441 aacatcaaca ctgcaatcaa cgttttaagc gacctcctgc ccgtgagaga atcaagtaag
     1501 gcagcaatat tggcgtgtgc cgctgaatac attcaaaaat tgaaagaaac cgatgaagca
     1561 aacatcgaaa agtggacgtt acaaaaattg cttagcgagc aaaacgcatc gcaattagcc
     1621 agtgcaaatg agaaactgca ggaagaactg ggaaatgctt acaaagaaat cgagtacatg
     1681 aaacgcgttt taaggaagga gggaatagaa tacgaggata tgcacactca caagaaacaa
     1741 gagaacgaaa gaaaaagcac taggagcgat aatccacatg aggctagagt gtcgcatagt
     1801 gatactaatt ctagcaaacc caaaaacacg aaggagaacc tttcaaaaag ctcttggagg
     1861 caagaatggc ttgcaaattt gaaacttatt tccgtttcgt tggttgatga gttcccttcg
     1921 gagctttccg acagtgatag acaaataatt aacgaaaaaa tgcagttact taaagatata
     1981 tttgctaaca accttaaatc agcaatttcc aataatttta gagagagtga catcattata
     2041 ctgaaaggtg aaatagaaga ttacccaatg agttctgaaa ttaagattta ctacaacgaa
     2101 ttacagaaca agcctgatgc aaaaaaagcc agattttggt catttatgaa gactcagaga
     2161 tttgtttcca acatgggatt tgatattcag aagtcctgtg aacctgtttc tatatccacg
     2221 tcggtcaaac cacatgtggt cgagcccgag catatggcag atgccaaaat tatgcctaag
     2281 gatatactgc aaattacaaa aaaaccgtta atggttaaaa atgtgaagcc ttcttctccg
     2341 ccagatgtga agtctttggt gcagctgagc acaatggaaa cgaaaaccct accagaaaag
     2401 aagcaattcg acagtatttt taattctaac aaagcaaaaa taatccctgg aaatgggaag
     2461 cacgcatcag aaaacatctc actctctttc tcaagacctg cgtcctacgg ctatttttct
     2521 gttggaaaaa gggttcgggg atcctctaga ggtgaacaaa agttgatttc tgaagaagat
     2581 ttgaacggtg aacaaaagct aatctccgag gaagacttga acggtgaaca aaaattaatc
     2641 tcagaagaag acttgaacgg atcctctaga gtcgactgaa tcgtagatac tgaaaaaccc
     2701 cgcaagttca cttcaactgt gcatcgtgca ccatctcaat ttctttcatt tatacatcgt
     2761 tttgccttct tttatgtaac tatactcctc taagtttcaa tcttggccat gtaacctctg
     2821 atctatagaa ttttttaaat gactagaatt aatgcccatc ttttttttgg acctaaattc
     2881 ttcatgaaaa tatattacga gggcttattc agaagctttg gacttcttcg ccagaggttt
     2941 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
     3001 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     3061 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     3121 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     3181 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     3241 gccgagcaaa tgcctgcaaa tcgctcccca tttcacccaa ttgtagatat gctaactcca
     3301 gcaatgagtt gatgaatctc ggtgtgtatt ttatgtcctc agaggacaac acctgttgta
     3361 atcgttcttc cacacggatc ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3421 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3481 ctgtgcggta tttcacaccg cataatcgct gggccattct catgaagaat atcttgaatt
     3541 tattgtcata ttactagttg gtgtggaagt cctaatatcg gtgatcaata tagtggttga
     3601 catgctggct agtcaacatt gagccttttg atcatgcaaa tatattacgg tattttacaa
     3661 tcaaatatca aacttaacta ttgactttat aacttattta ggtggtaaca ttcttataaa
     3721 aaagaaaaaa attactgcaa aacagtacta gcttttaact tgtatcctag gttatctatg
     3781 ctgtctcacc atagagaata ttacctattt cagaatgtat gtccatgatt cgccgggtaa
     3841 atacatataa tacacaaatc tggcttaata aagtctataa tatatctcat aaagaagtgc
     3901 taaattggct agtgctatat atttttaaga aaatttcttt tgactaagtc catatcgact
     3961 ttgtaaaagt tcacattagc atacatatat tacacgagcc agaaatagta acttttgcct
     4021 aaatcacaaa ttgcaaaatt taattgcttg caaaaggtca catgcttata atcaactttt
     4081 ttaaaaattt aaaatacttt tttatttttt atttttaaac ataaatgaaa taatttattt
     4141 attgtttatg attaccgaaa cataaaacct gctcaagaaa aagaaactgt tttgtccttg
     4201 gaaaaaaagc actacctagg agcggccaaa atgccgaggc tttcatagct taaactcttt
     4261 acagaaaata ggcattatag atcagttcga gttttcttat tcttccttcc ggttttatcg
     4321 tcacagtttt acagtaaata agtatcacct cttagagttc gatgataagc tgtcaaacat
     4381 gagaattaat tccacatgtt aaaatagtga aggagcatgt tcggcacaca gtggaccgaa
     4441 cgtggggtaa gtgcactagg gtccggttaa acggatctcg cattgatgag gcaacgctaa
     4501 ttatcaacat atagattgtt atctatctgc atgaacacga aatctttact tgacgacttg
     4561 aggctgatgg tgtttatgca aagaaaccac tgtgtttaat atgtgtcact gtttgatatt
     4621 actgtcagcg tagaagataa tagtaaaagc ggttaataag tgtatttgag ataagtgtga
     4681 taaagttttt acagcgaaaa gacgataaat acaagaaaat gattacgagg atacggagag
     4741 aggtatgtac atgtgtattt atatactaag ctgccggcgg ttgtttgcaa gaccgagaaa
     4801 aggctagcaa gaatcgggtc attgtagcgt atgcgcctgt gaacattctc ttcaacaagt
     4861 ttgattccat tgcggtgaaa tggtaaaagt caaccccctg cgatgtatat tttcctgtac
     4921 aatcaatcaa aaagccaaat gatttagcat tatctttaca tcttgttatt ttacagattt
     4981 tatgtttaga tcttttatgc ttgcttttca aaaggcttgc aaggccgttt ctgacagagt
     5041 aaaattcttg agggaacttt caccattatg ggaaatggtt caagaaggta ttgacttaaa
     5101 ctccatcaaa tggtcaggtc attgagtgtt ttttatttgt tgtatttttt tttttttaga
     5161 gaaaatcctc caatatataa attaggaatc atagtttcat gattttctgt tacacctaac
     5221 tttttgtgtg gtgccctcct ccttgtcaat attaatgtta aagtgcaatt ctttttcctt
     5281 atcacgttga gccattagta tcaatttgct tacctgtatt cctttacatc ctcctttttc
     5341 tccttcttga taaatgtatg tagattgcgt atatagtttc gtctacccta tgaacatatt
     5401 ccattttgta atttcgtgtc gtttctatta tgaatttcat ttataaagtt tatgtacaaa
     5461 tatcataaaa aaagagaatc tttttaagca aggattttct taacttcttc ggcgacagca
     5521 tcaccgactt cggtggtact gttggaacca cctaaatcac cagttctgat acctgcatcc
     5581 aaaacctttt taactgcatc ttcaatggcc ttaccttctt caggcaagtt caatgacaat
     5641 ttcaacatca ttgcagcaga caagatagtg gcgatagggt tgaccttatt ctttggcaaa
     5701 tctggagcag aaccgtggca tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa
     5761 gaggccaagg acgcagatgg caacaaaccc aaggaacctg ggataacgga ggcttcatcg
     5821 gagatgatat caccaaacat gttgctggtg attataatac catttaggtg ggttgggttc
     5881 ttaactagga tcatggcggc agaatcaatc aattgatgtt gaaccttcaa tgtagggaat
     5941 tcgttcttga tggtttcctc cacagttttt ctccataatc ttgaagaggc caaaacatta
     6001 gctttatcca aggaccaaat aggcaatggt ggctcatgtt gtagggccat gaaagcggcc
     6061 attcttgtga ttctttgcac ttctggaacg gtgtattgtt cactatccca agcgacacca
     6121 tcaccatcgt cttcctttct cttaccaaag taaatacctc ccactaattc tctgacaaca
     6181 acgaagtcag tacctttagc aaattgtggc ttgattggag ataagtctaa aagagagtcg
     6241 gatgcaaagt tacatggtct taagttggcg tacaattgaa gttctttacg gatttttagt
     6301 aaaccttgtt caggtctaac actaccggta ccccatttag gaccacccac agcacctaac
     6361 aaaacggcat cagccttctt ggaggcttcc agcgcctcat ctggaagtgg aacacctgta
     6421 gcatcgatag cagcaccacc aattaaatga ttttcgaaat cgaacttgac attggaacga
     6481 acatcagaaa tagctttaag aaccttaatg gcttcggctg tgatttcttg accaacgtgg
     6541 tcacctggca aaacgacgat cttcttaggg gcagacatta gaatggtata tccttgaaat
     6601 atatatatat atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc
     6661 taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt
     6721 gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcgct
     6781 ctcacctttc ctttttctcc caatttttca gttgaaaaag gtatatgcgt caggcgacct
     6841 ctgaaattaa caaaaaattt ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg
     6901 tgttctcgtt atgttgagga aaaaaataat ggttgctaag agattcgaac tcttgcatct
     6961 tacgatacct gagtattccc acagttaact gcggtcaaga tatttcttga atcaggcgcc
     7021 ttagaccgct cggccaaaca accaattact tgttgagaaa tagagtataa ttatcctata
     7081 aatataacgt ttttgaacac acatgaacaa ggaagtacag gacaattgat tttgaagaga
     7141 atgtggattt tgatgtaatt gttgggattc catttttaat aaggcaataa tattaggtat
     7201 gtagatatac tagaagttct cctcgaggaa ttcttgaaga cgaaagggcc tcgtgatacg
     7261 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     7321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     7381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     7441 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     7501 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     7561 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     7621 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     7681 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     7741 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     7801 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     7861 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     7921 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     7981 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     8041 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     8101 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     8161 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     8221 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     8281 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     8341 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     8401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     8461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     8521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     8581 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     8641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     8701 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     8761 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8821 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8881 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8941 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     9001 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     9061 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     9121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     9241 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatccg
     9301 ggatcgaaga aatgatggta aatgaaatag gaaatcaagg agcatgaagg caaaagacaa
     9361 atataagggt cgaacgaaaa ataaagtgaa aagtgttgat atgatgtatt tggctttgcg
     9421 gcgccgaaaa aacgagttta cgcaattgca caatcatgct gactctgtgg cggacccgcg
     9481 ctcttgccgg cccggcgata acgctgggcg tgaggctgtg cccggcggag ttttttgcgc
     9541 ctgcattttc caaggtttac cctgcgctaa ggggcgagat tggagaagca ataagaatgc
     9601 cggttggggt tgcgatgatg acgaccacga caactggtgt cattatttaa gttgccgaaa
     9661 gaacctgagt gcatttgcaa catgagtata ctagaagaat gagccaagac ttgcgagacg
     9721 cgagtttgcc ggtggtgcga acaatagagc gaccatgacc ttgaaggtga gacgcgcata
     9781 accgctagag tactttgaag aggaaacagc aatagggttg ctaccagtat aaatagacag
     9841 gtacatacaa cactggaaat ggttgtctgt ttgagtacgc tttcaattca tttgggtgtg
     9901 cactttatta tgttacaata tggaagggaa ctttacactt ctcctatgca catatattaa
     9961 ttaaagtcca atgctagtag agaagggggg taacacccct ccgcgctctt ttccgatttt
    10021 tttctaaacc gtggaatatt tcggat
//
