LOCUS       sequence-381349-        9886 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 9886)
  AUTHORS   Recio PS, Mitra NJ, Shively CA, Song D, Jaramillo G, Lewis KS, Chen 
            X, Mitra RD
  TITLE     Zinc cluster transcription factors frequently activate target genes 
            using a non-canonical half-site binding mode.
  JOURNAL   Nucleic Acids Res. 2023 Jun 9;51(10):5006-5021. doi: 
            10.1093/nar/gkad320.
  PUBMED    37125648
REFERENCE   2  (bases 1 to 9886)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2023-06-9- 9"; volume: "51"; issue: 
            "10"; pages: "5006-5021"
COMMENT     Sequence Label: ADH1pro_TEA1_SIR4
FEATURES             Location/Qualifiers
     source          1..9886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3581..3610
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3617..3646
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3653..3682
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4077..4264
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4132..4156)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    5094..6256
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      5419..6256
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(6095..6769)
                     /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(6770..6871)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6891..6909)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6977..7081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7082..7150,7151..7942)
                     /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(7300..7319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8113..8701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8602..8621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8855..8872
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(9690..9886,1..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 gaaagatgac
      541 agcgccattg tggccaaaca agaatgaaaa aaaccacaca gttaaaagag cgttatcaac
      601 ggatatgacc agcaatattt tgagtagcac aaacgcgagc tcaaacgaag aaaattctag
      661 gagttcttct gcagccaatg tacgctccgg aacaggtgca aatacactta ctaatggcgg
      721 cagtactaga aagagacttg cgtgcactaa ttgtagaaat agaaggaaaa aatgtgattt
      781 aggatttccc tgtggtaact gttccaggtt ggaattggtt tgtaatgtta atgacgagga
      841 cttaagaaag aagcggtaca ctaataaata tgtcaagtct ttggagagcc atattgctca
      901 actggagacc aacttaaaaa acctagttca gaagatctac cctgacgatg agcaaatact
      961 gaaccgaatg atggtaggtg atgtattatc agctctacca gacagttcac aagtctcaat
     1021 caattatact gaccaaactc cctctcttcc aattcccgca accagaggta cattcattat
     1081 tgaaaacgat aaggtcagtc aacctctatc gtcctttaac caacaaacag agccatctac
     1141 tctaaactcg ggtatcttca acacccaaaa acaaaatttc gaagaatccc ttgatgatca
     1201 gttactttta cgaagatcgt taacaccgca aggtgaaaaa aagaagaaac cgttggtaaa
     1261 aggtagtctt tatcctgaag gacctgtcag ttacaaacgg aagcaccccg taaaatcgga
     1321 cagtttattg cctgtgtctt cgttaacagc tgctacagac ccatctactt tttctgacgg
     1381 tataactgct ggtaattccg tcctagttaa tggtgaactg aaaaaacgta tatccgactt
     1441 gaagaccacc gtaatagtaa gaggactaaa cgatgataat cccaactcta tcaataacga
     1501 tcccaggatt ttaaaatctc tttccaattt ctataagtgg ctgtatccag gttattttat
     1561 ttttgttcac agagaaagtt ttctttatgg attcttcaat cactccaaga ataactatga
     1621 agactcaagt tactgttctg tcgaattgat atatgccatg tgcgctgtcg gttcgaggct
     1681 tacaccggat ctacaagaat attcagaagt gtattatcaa cgaagtaaga agactttatt
     1741 acagctcgtc tttgatgagc aaagtacagc ccgtatcacc accgtccaag cgctattttg
     1801 cttagccttt tacgaactgg ggaagggtaa taatcaatta gggtggtact tctcgggcct
     1861 ggctatcagg gtcggctacg acatgggatt tcaactggac cctaaagtct ggtacgttga
     1921 tgataacaat ctgcagctga cccaaagtga gttggaaata agatcaagga tttactgggg
     1981 ttgctatata gcggatcatt ttatttgcct gatgttgggt agaacgtcca cattaagcgt
     2041 tagtaactca accatgccag agtcggatga gctgccagaa gtcaacggca ccgaagagtt
     2101 cagatttatt ggaagacatg ttttacagat atccttgcct ctgaaaaact tgataattct
     2161 atcaaggctt gtacaaatct tcacctctaa aattttcatc gagtcggaag atatagcacg
     2221 aaaactaaag tacttgaaca ctttcaattc tcaggtttac aactggagac aatctctacc
     2281 ggaatttttg caatggtcga aaacactcat cgagaacgat gacgtatcca cagaccccac
     2341 tatatcttac ttctggtatt gctactacat agtgcggctc acctttaaca agccatttat
     2401 agaagactct caagaatccg aaacggtagt catagagatc atcgatgatt tgaagacttt
     2461 gctggacaat ttcgggaaaa aattcggaaa ctacacgaag ggaaaccttt accaacttta
     2521 ctcatgcctg ttggccatca actgtttgaa gaagttgaaa gagatacgtt ccagtgagca
     2581 agactcatgg aatgcacaat tggatttttt caatcacatc ttctacacac aactatatcc
     2641 cgcatatgat ctgccgaaaa agctgcagga ggatacggag ctggaaacag aacaagaaaa
     2701 ccagatgttg aaccaagttg gaaatataaa ctacacgcac gatttttcac tctcccacga
     2761 aattgacgat cttatcagag aactgtttgg ggtgggcaca cctcaaaaac tcagagtgtc
     2821 gcatagtgat actaattcta gcaaacccga aaacacgaag gagaaccttt caaaaagctc
     2881 ttggaggcag gaatggcttg caaatttgaa acttatttcc gtttcgttgg ttgatgagtt
     2941 cccttcggag ctttccgaca gtgatagaca agtaattaac gaaaaaatgc agttacttaa
     3001 agatatattt gctaacaacc ttaaatcagc aatttccaat aattttagag agagtgacat
     3061 cattatactg aaaggtgaaa tagaagatta cccaatgagt tctgaaatta agatttacta
     3121 caacgaatta cagaacaagc ctgatgcaaa aaaagccaga ttttggtcat ttatgaagac
     3181 tcagagattt gtttccaaca tgggatttga tattcggaag tcctgtgaac ctgtttctat
     3241 atccacgtcg gtcaaaccac atgtggtcga gcccgagcat atggcagatg ccaaaattat
     3301 gcctaaggat gtactgcaaa ttacaaaaaa accgttaatg gttaaaaatg tgaagccttc
     3361 ttctccgcca gatgtgaagt ctttggtgca gctgagcaca atggaaacga aaaccctacc
     3421 agaaaagaag caattcgaca gtatttttaa ttctaacaaa gcaaaaataa tccctggaaa
     3481 tgggaagcac gcatcagaaa acatctcact ctctttctca agacctgcgt cctacggcta
     3541 tttttctgtt ggaaaaaggg ttcggggatc ctctagaggt gaacaaaagt tgatttctga
     3601 agaagatttg aacggtgaac aaaagctaat ctccgaggaa gacttgaacg gtgaacaaaa
     3661 attaatctca gaagaagact tgaacggatc ctctagagtc gactgaatcg tagatactga
     3721 aaaaccccgc aagttcactt caactgtgca tcgtgcacca tctcaatttc tttcatttat
     3781 acatcgtttt gccttctttt atgtaactat actcctctaa gtttcaatct tggccatgta
     3841 acctctgatc tatagaattt tttaaatgac tagaattaat gcccatcttt tttttggacc
     3901 taaattcttc atgaaaatat attacgaggg cttattcaga agctttggac ttcttcgcca
     3961 gaggtttggt caagtctcca atcaaggttg tcggcttgtc taccttgcca gaaatttacg
     4021 aaaagatgga aaagggtcaa atcgttggta gatacgttgt tgacacttct aaataagcga
     4081 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     4141 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     4201 tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta
     4261 ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt cacccaattg tagatatgct
     4321 aactccagca atgagttgat gaatctcggt gtgtatttta tgtcctcaga ggacaacacc
     4381 tgttgtaatc gttcttccac acggatcctg gcgtaatagc gaagaggccc gcaccgatcg
     4441 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     4501 tacgcatctg tgcggtattt cacaccgcat aatcgctggg ccattctcat gaagaatatc
     4561 ttgaatttat tgtcatatta ctagttggtg tggaagtcct aatatcggtg atcaatatag
     4621 tggttgacat gctggctagt caacattgag ccttttgatc atgcaaatat attacggtat
     4681 tttacaatca aatatcaaac ttaactattg actttataac ttatttaggt ggtaacattc
     4741 ttataaaaaa gaaaaaaatt actgcaaaac agtactagct tttaacttgt atcctaggtt
     4801 atctatgctg tctcaccata gagaatatta cctatttcag aatgtatgtc catgattcgc
     4861 cgggtaaata catataatac acaaatctgg cttaataaag tctataatat atctcataaa
     4921 gaagtgctaa attggctagt gctatatatt tttaagaaaa tttcttttga ctaagtccat
     4981 atcgactttg taaaagttca cattagcata catatattac acgagccaga aatagtaact
     5041 tttgcctaaa tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat gcttataatc
     5101 aactttttta aaaatttaaa atactttttt attttttatt tttaaacata aatgaaataa
     5161 tttatttatt gtttatgatt accgaaacat aaaacctgct caagaaaaag aaactgtttt
     5221 gtccttggaa aaaaagcact acctaggagc ggccaaaatg ccgaggcttt catagcttaa
     5281 actctttaca gaaaataggc attatagatc agttcgagtt ttcttattct tccttccggt
     5341 tttatcgtca cagttttaca gtaaataagt atcacctctt agagttcgat gataagctgt
     5401 caaacatgag aattaattcc acatgttaaa atagtgaagg agcatgttcg gcacacagtg
     5461 gaccgaacgt ggggtaagtg cactagggtc cggttaaacg gatctcgcat tgatgaggca
     5521 acgctaatta tcaacatata gattgttatc tatctgcatg aacacgaaat ctttacttga
     5581 cgacttgagg ctgatggtgt ttatgcaaag aaaccactgt gtttaatatg tgtcactgtt
     5641 tgatattact gtcagcgtag aagataatag taaaagcggt taataagtgt atttgagata
     5701 agtgtgataa agtttttaca gcgaaaagac gataaataca agaaaatgat tacgaggata
     5761 cggagagagg tatgtacatg tgtatttata tactaagctg ccggcggttg tttgcaagac
     5821 cgagaaaagg ctagcaagaa tcgggtcatt gtagcgtatg cgcctgtgaa cattctcttc
     5881 aacaagtttg attccattgc ggtgaaatgg taaaagtcaa ccccctgcga tgtatatttt
     5941 cctgtacaat caatcaaaaa gccaaatgat ttagcattat ctttacatct tgttatttta
     6001 cagattttat gtttagatct tttatgcttg cttttcaaaa ggcttgcagg caagtgcaca
     6061 aacaatactt aaataaatac tactcagtaa taacctattt cttagcattt ttgacgaaat
     6121 ttgctatttt gttagagtct tttacaccat ttgtctccac acctccgctt acatcaacac
     6181 caataacgcc atttaatcta agcgcatcac caacattttc tggcgtcagt ccaccagcta
     6241 acataaaatg taagctctcg gggctctctt gccttccaac ccagtcagaa atcgagttcc
     6301 aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa caagggaata aacgaatgag
     6361 gtttctgtga agctgcactg agtagtatgt tgcagtcttt tggaaatacg agtcttttaa
     6421 taactggcaa accgaggaac tcttggtatt cttgccacga ctcatctcca tgcagttgga
     6481 cgatatcaat gccgtaatca ttgaccagag ccaaaacatc ctccttaggt tgattacgaa
     6541 acacgccaac caagtatttc ggagtgcctg aactattttt atatgctttt acaagacttg
     6601 aaattttcct tgcaataacc gggtcaattg ttctctttct attgggcaca catataatac
     6661 ccagcaagtc agcatcggaa tctagtgcac attctgcggc ctctgtgctc tgcaagccgc
     6721 aaactttcac caatggacca gaactacctg tgaaattaat aacagacata ctccaagctg
     6781 cctttgtgtg cttaatcacg tatactcacg tgctcaatag tcaccaatgc cctccctctt
     6841 ggccctctcc ttttcttttt tcgaccgaat taattcttga agacgaaagg gcctcgtgat
     6901 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6961 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7321 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8761 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8821 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8881 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat
     8941 ccgggatcga agaaatgatg gtaaatgaaa taggaaatca aggagcatga aggcaaaaga
     9001 caaatataag ggtcgaacga aaaataaagt gaaaagtgtt gatatgatgt atttggcttt
     9061 gcggcgccga aaaaacgagt ttacgcaatt gcacaatcat gctgactctg tggcggaccc
     9121 gcgctcttgc cggcccggcg ataacgctgg gcgtgaggct gtgcccggcg gagttttttg
     9181 cgcctgcatt ttccaaggtt taccctgcgc taaggggcga gattggagaa gcaataagaa
     9241 tgccggttgg ggttgcgatg atgacgacca cgacaactgg tgtcattatt taagttgccg
     9301 aaagaacctg agtgcatttg caacatgagt atactagaag aatgagccaa gacttgcgag
     9361 acgcgagttt gccggtggtg cgaacaatag agcgaccatg accttgaagg tgagacgcgc
     9421 ataaccgcta gagtactttg aagaggaaac agcaataggg ttgctaccag tataaataga
     9481 caggtacata caacactgga aatggttgtc tgtttgagta cgctttcaat tcatttgggt
     9541 gtgcacttta ttatgttaca atatggaagg gaactttaca cttctcctat gcacatatat
     9601 taattaaagt ccaatgctag tagagaaggg gggtaacacc cctccgcgct cttttccgat
     9661 ttttttctaa accgtggaat atttcggata tccttttgtt gtttccgggt gtacaatatg
     9721 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     9781 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     9841 ataatgggct aaacaagact acaccaatta cactgcctca ttgatg
//
