LOCUS       sequence-334951-        9881 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 9881)
  TITLE     Mitra lab plasmids
REFERENCE   2  (bases 1 to 9881)
  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_GCN4_SIR4
FEATURES             Location/Qualifiers
     source          1..9881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(481..499)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        567..671
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(672..740,741..1532)
                     /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(890..909)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1703..2291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2192..2211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2445..2462
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3280..3984
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4783..4839
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5668..5697
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5704..5733
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5740..5769
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      6164..6351
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6219..6243)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    7181..7473
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     CDS             complement(8598..9692)
                     /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(join(9693..9881,1..219))
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 cggttccggc gctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg
       61 cgtcaggcga cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga
      121 tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg
      181 aactcttgca tcttacgata cctgagtatt cccacagtta actgcggtca agatatttct
      241 tgaatcaggc gccttagacc gctcggccaa acaaccaatt acttgttgag aaatagagta
      301 taattatcct ataaatataa cgtttttgaa cacacatgaa caaggaagta caggacaatt
      361 gattttgaag agaatgtgga ttttgatgta attgttggga ttccattttt aataaggcaa
      421 taatattagg tatgtagata tactagaagt tctcctcgag gaattcttga agacgaaagg
      481 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      541 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      601 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      661 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      721 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      781 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      841 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      901 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      961 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1021 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1081 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1141 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1201 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1261 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1321 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1381 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1441 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1501 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1561 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1621 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1681 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1741 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1801 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1861 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1921 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1981 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2041 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2101 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2161 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2221 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2281 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2341 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2401 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2461 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2521 aatgcaggat ccgggatcga agaaatgatg gtaaatgaaa taggaaatca aggagcatga
     2581 aggcaaaaga caaatataag ggtcgaacga aaaataaagt gaaaagtgtt gatatgatgt
     2641 atttggcttt gcggcgccga aaaaacgagt ttacgcaatt gcacaatcat gctgactctg
     2701 tggcggaccc gcgctcttgc cggcccggcg ataacgctgg gcgtgaggct gtgcccggcg
     2761 gagttttttg cgcctgcatt ttccaaggtt taccctgcgc taaggggcga gattggagaa
     2821 gcaataagaa tgccggttgg ggttgcgatg atgacgacca cgacaactgg tgtcattatt
     2881 taagttgccg aaagaacctg agtgcatttg caacatgagt atactagaag aatgagccaa
     2941 gacttgcgag acgcgagttt gccggtggtg cgaacaatag agcgaccatg accttgaagg
     3001 tgagacgcgc ataaccgcta gagtactttg aagaggaaac agcaataggg ttgctaccag
     3061 tataaataga caggtacata caacactgga aatggttgtc tgtttgagta cgctttcaat
     3121 tcatttgggt gtgcacttta ttatgttaca atatggaagg gaactttaca cttctcctat
     3181 gcacatatat taattaaagt ccaatgctag tagagaaggg gggtaacacc cctccgcgct
     3241 cttttccgat ttttttctaa accgtggaat atttcggata tccttttgtt gtttccgggt
     3301 gtacaatatg gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata
     3361 ccttcgttgg tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac
     3421 cagacaagac ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg
     3481 tacataacga actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca
     3541 ctaccctttt tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct
     3601 ttttttttct tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat
     3661 gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt
     3721 ccagagctga tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca
     3781 cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata
     3841 aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt
     3901 ttcctcgtca ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatacttcaa
     3961 gctataccaa gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgtccgaa
     4021 tatcagccaa gtttatttgc tttaaatcca atgggtttct caccattgga tggttctaaa
     4081 tcaaccaacg aaaatgtatc tgcttccact tctactgcca aaccaatggt tggccaattg
     4141 atttttgata aattcatcaa gactgaagag gatccaatta tcaaacagga taccccttcg
     4201 aaccttgatt ttgattttgc tcttccacaa acggcaactg cacctgatgc caagaccgtt
     4261 ttgccaattc cggagctaga tgacgctgta gtggaatctt tcttttcgtc aagcactgat
     4321 tcaactccaa tgtttgagta tgaaaaccta gaagacaact ctaaagaatg gacatccttg
     4381 tttgacaatg acattccagt taccactgac gatgtttcat tggctgataa ggcaattgaa
     4441 tccactgaag aagtttctct ggtaccatcc aatctggaag tctcgacaac ttcattctta
     4501 cccactcctg ttctagaaga tgctaaactg actcaaacaa gaaaggttaa gaaaccaaat
     4561 tcagtcgtta agaagtcaca tcatgttgga aaggatgacg aatcgagact ggatcatcta
     4621 ggtgttgttg cttacaaccg caaacagcgt tcgattccac tttctccaat tgtgcccgaa
     4681 tccagtgatc ctgctgctct aaaacgtgct agaaacactg aagccgccag gcgttctcgt
     4741 gcgagaaagt tgcaaagaat gaaacaactt gaagacaagg ttgaagaatt gctttcgaaa
     4801 aattatcact tggaaaatga ggttgccaga ttaaagaaat tagttggcga acgccgtacg
     4861 ctgcaggtcg acggtggcgg ttctggcggt ggcggatcca gagtgtcgca tagtgatact
     4921 aattctagca aacccaaaaa cacgaaggag aacctttcaa aaagctcttg gaggcaagaa
     4981 tggcttgcaa atttgaaact tatttccgtt tcgttggttg atgagttccc ttcggagctt
     5041 tccgacagtg atagacaaat aattaacgaa aaaatgcagt tacttaaaga tatatttgct
     5101 aacaacctta aatcagcaat ttccaataat tttagagaga gtgacatcat tatactgaaa
     5161 ggtgaaatag aagattaccc aatgagttct gaaattaaga tttactacaa cgaattacag
     5221 aacaagcctg atgcaaaaaa agccagattt tggtcattta tgaagactca gagatttgtt
     5281 tccaacatgg gatttgatat tcagaagtcc tgtgaacctg tttctatatc cacgtcggtc
     5341 aaaccacatg tggtcgagcc cgagcatatg gcagatgcca aaattatgcc taaggatata
     5401 ctgcaaatta caaaaaaacc gttaatggtt aaaaatgtga agccttcttc tccgccagat
     5461 gtgaagtctt tggtgcagct gagcacaatg gaaacgaaaa ccctaccaga aaagaagcaa
     5521 ttcgacagta tttttaattc taacaaagca aaaataatcc ctggaaatgg gaagcacgca
     5581 tcagaaaaca tctcactctc tttctcaaga cctgcgtcct acggctattt ttctgttgga
     5641 aaaagggttc ggggatcctc tagaggtgaa caaaagttga tttctgaaga agatttgaac
     5701 ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa
     5761 gaagacttga acggatcctc tagagtcgac tgaatcgtag atactgaaaa accccgcaag
     5821 ttcacttcaa ctgtgcatcg tgcaccatct caatttcttt catttataca tcgttttgcc
     5881 ttcttttatg taactatact cctctaagtt tcaatcttgg ccatgtaacc tctgatctat
     5941 agaatttttt aaatgactag aattaatgcc catctttttt ttggacctaa attcttcatg
     6001 aaaatatatt acgagggctt attcagaagc tttggacttc ttcgccagag gtttggtcaa
     6061 gtctccaatc aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa
     6121 gggtcaaatc gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt
     6181 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     6241 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     6301 gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg gcatgccgag
     6361 caaatgcctg caaatcgctc cccatttcac ccaattgtag atatgctaac tccagcaatg
     6421 agttgatgaa tctcggtgtg tattttatgt cctcagagga caacacctgt tgtaatcgtt
     6481 cttccacacg gatcctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6541 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     6601 ggtatttcac accgcataat cgctgggcca ttctcatgaa gaatatcttg aatttattgt
     6661 catattacta gttggtgtgg aagtcctaat atcggtgatc aatatagtgg ttgacatgct
     6721 ggctagtcaa cattgagcct tttgatcatg caaatatatt acggtatttt acaatcaaat
     6781 atcaaactta actattgact ttataactta tttaggtggt aacattctta taaaaaagaa
     6841 aaaaattact gcaaaacagt actagctttt aacttgtatc ctaggttatc tatgctgtct
     6901 caccatagag aatattacct atttcagaat gtatgtccat gattcgccgg gtaaatacat
     6961 ataatacaca aatctggctt aataaagtct ataatatatc tcataaagaa gtgctaaatt
     7021 ggctagtgct atatattttt aagaaaattt cttttgacta agtccatatc gactttgtaa
     7081 aagttcacat tagcatacat atattacacg agccagaaat agtaactttt gcctaaatca
     7141 caaattgcaa aatttaattg cttgcaaaag gtcacatgct tataatcaac ttttttaaaa
     7201 atttaaaata cttttttatt ttttattttt aaacataaat gaaataattt atttattgtt
     7261 tatgattacc gaaacataaa acctgctcaa gaaaaagaaa ctgttttgtc cttggaaaaa
     7321 aagcactacc taggagcggc caaaatgccg aggctttcat agcttaaact ctttacagaa
     7381 aataggcatt atagatcagt tcgagttttc ttattcttcc ttccggtttt atcgtcacag
     7441 ttttacagta aataagtatc acctcttaga gttcgatgat aagctgtcaa acatgagaat
     7501 taattccaca tgttaaaata gtgaaggagc atgttcggca cacagtggac cgaacgtggg
     7561 gtaagtgcac tagggtccgg ttaaacggat ctcgcattga tgaggcaacg ctaattatca
     7621 acatatagat tgttatctat ctgcatgaac acgaaatctt tacttgacga cttgaggctg
     7681 atggtgttta tgcaaagaaa ccactgtgtt taatatgtgt cactgtttga tattactgtc
     7741 agcgtagaag ataatagtaa aagcggttaa taagtgtatt tgagataagt gtgataaagt
     7801 ttttacagcg aaaagacgat aaatacaaga aaatgattac gaggatacgg agagaggtat
     7861 gtacatgtgt atttatatac taagctgccg gcggttgttt gcaagaccga gaaaaggcta
     7921 gcaagaatcg ggtcattgta gcgtatgcgc ctgtgaacat tctcttcaac aagtttgatt
     7981 ccattgcggt gaaatggtaa aagtcaaccc cctgcgatgt atattttcct gtacaatcaa
     8041 tcaaaaagcc aaatgattta gcattatctt tacatcttgt tattttacag attttatgtt
     8101 tagatctttt atgcttgctt ttcaaaaggc ttgcaaggcc gtttctgaca gagtaaaatt
     8161 cttgagggaa ctttcaccat tatgggaaat ggttcaagaa ggtattgact taaactccat
     8221 caaatggtca ggtcattgag tgttttttat ttgttgtatt tttttttttt tagagaaaat
     8281 cctccaatat ataaattagg aatcatagtt tcatgatttt ctgttacacc taactttttg
     8341 tgtggtgccc tcctccttgt caatattaat gttaaagtgc aattcttttt ccttatcacg
     8401 ttgagccatt agtatcaatt tgcttacctg tattccttta catcctcctt tttctccttc
     8461 ttgataaatg tatgtagatt gcgtatatag tttcgtctac cctatgaaca tattccattt
     8521 tgtaatttcg tgtcgtttct attatgaatt tcatttataa agtttatgta caaatatcat
     8581 aaaaaaagag aatcttttta agcaaggatt ttcttaactt cttcggcgac agcatcaccg
     8641 acttcggtgg tactgttgga accacctaaa tcaccagttc tgatacctgc atccaaaacc
     8701 tttttaactg catcttcaat ggccttacct tcttcaggca agttcaatga caatttcaac
     8761 atcattgcag cagacaagat agtggcgata gggttgacct tattctttgg caaatctgga
     8821 gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt tcttgtctgg caaagaggcc
     8881 aaggacgcag atggcaacaa acccaaggaa cctgggataa cggaggcttc atcggagatg
     8941 atatcaccaa acatgttgct ggtgattata ataccattta ggtgggttgg gttcttaact
     9001 aggatcatgg cggcagaatc aatcaattga tgttgaacct tcaatgtagg gaattcgttc
     9061 ttgatggttt cctccacagt ttttctccat aatcttgaag aggccaaaac attagcttta
     9121 tccaaggacc aaataggcaa tggtggctca tgttgtaggg ccatgaaagc ggccattctt
     9181 gtgattcttt gcacttctgg aacggtgtat tgttcactat cccaagcgac accatcacca
     9241 tcgtcttcct ttctcttacc aaagtaaata cctcccacta attctctgac aacaacgaag
     9301 tcagtacctt tagcaaattg tggcttgatt ggagataagt ctaaaagaga gtcggatgca
     9361 aagttacatg gtcttaagtt ggcgtacaat tgaagttctt tacggatttt tagtaaacct
     9421 tgttcaggtc taacactacc ggtaccccat ttaggaccac ccacagcacc taacaaaacg
     9481 gcatcagcct tcttggaggc ttccagcgcc tcatctggaa gtggaacacc tgtagcatcg
     9541 atagcagcac caccaattaa atgattttcg aaatcgaact tgacattgga acgaacatca
     9601 gaaatagctt taagaacctt aatggcttcg gctgtgattt cttgaccaac gtggtcacct
     9661 ggcaaaacga cgatcttctt aggggcagac attagaatgg tatatccttg aaatatatat
     9721 atatatattg ctgaaatgta aaaggtaaga aaagttagaa agtaagacga ttgctaacca
     9781 cctattggaa aaaacaatag gtccttaaat aatattgtca acttcaagta ttgtgatgca
     9841 agcatttagt catgaacgct tctctattct atatgaaaag c
//
