LOCUS       sequence-381347-       10108 bp    DNA     circular SYN 15-APR-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 10108)
  TITLE     A Novel Mode of Half Site Binding in Budding Yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: ADH1pro_YRR1_SIR4
FEATURES             Location/Qualifiers
     source          1..10108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    545..569
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3006..3030)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3803..3832
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3839..3868
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3875..3904
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4299..4486
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4354..4378)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    5316..6478
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      5641..6478
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(6317..6991)
                     /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(6992..7093)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7113..7131)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7199..7303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7304..7372,7373..8164)
                     /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(7522..7541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8335..8923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8824..8843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9077..9094
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(9912..10108,1..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 gaaagggcgc
      541 gccaacaagt ttgtacaaaa aagcaggcta caaaatgaaa agaagaagcg atgctttgtt
      601 gggaagtttc caggccacca acgttacgcc gccaagcgat aatagtaata gtacggccgg
      661 tggagccaat ggaagtaact caggaacacc aacttctact agcggtaaga agagaaacaa
      721 gcttataaaa tcttgtggct tttgcagaag gagaaaactt cgttgtgatc agcaaaaacc
      781 tatgtgttct acatgcattt ctagaaactt aacaacctgt caatatgctg aagaattcaa
      841 caaaaacatc gaaaagaaag ctacctatgg tccctatcct aacgccgatt tacttaagaa
      901 agttgaagaa ctagaaaaca aaatacgtat tctagaagct gaaaaaaaca ctaattcgtc
      961 tgcgagctcc atgtacactt cgccaaattt ccctccttta ggcactagtg taggtagagg
     1021 ttctactgaa acttcatctc cattacccga tggtgtaata aatccatatg ccgaccggta
     1081 ctacctacaa agtaaacatt ccggaagatc aacactatac ggccccactt ctatgagaac
     1141 ccaaattgca aatagtaatt gggggttcat tgaaaaatat aaacaattat gggcgaaggt
     1201 taaagtagaa agaaataaat ggaagcaaaa caaccaaaaa acgatgtgca gggaactagg
     1261 ccttttggat gagtcggatt ggcagccaga tccattaatc aaacagatat gtcgtttcct
     1321 accatcatat aacaaagctt tgtctatttt agatgatttc tttaatgatg gagcatgcaa
     1381 tgagatcaac gtgattcttg ataaggcaaa ggttagaaga gactttttag attatttcat
     1441 gcccgaaaaa gaggtaaagg cggaaggtga cagatccata gtttatattt tatccaatcc
     1501 aaagaaaaat tattacaaag ctgctgtgat actattaatc ctatgtctga aatacttcca
     1561 tacagatgtc ccaaccccaa ttgagaagtt ctttacttta ctaaaggggg cctccaccgc
     1621 taaggtgttc tacattgagc gtgcacaaat gttaatactt ttctactacc atcgtgaaac
     1681 atattcattc ggtggagatg gctccgattt ggttaatata aatgaatgtt tagttactac
     1741 ggttactaca atagggttgc atttaaacat tagagaaact ttcaaggaac atgaggtttt
     1801 tatgggtagc atcgaaagtt tagaaaatgt ctggttaatg gcaatattta tcgattataa
     1861 catatcttgt aacgttggta gacccttatt gattaacaaa ttttatctag acgaaaacca
     1921 agatcattgc atattgaata gtaaaagtaa aacatatgag gggaagctga aaagatattt
     1981 aaaacttaca aggccaatgt tgttaactct ttatgataga gataaattcc cggatttgaa
     2041 agcttattcc aagcgaatta tcaattttgt agaagaagaa cttggtccac ttggacatta
     2101 cacaggtgaa aatatatcgg aagaagttcc attaagggaa agtagaattt taagtatggc
     2161 agtcggacta ttattgtcgt tttatgcact aatacattct gtgctgaaag tcagaaacat
     2221 agaatccaaa aataatacgt ttcaattggt tttgatcaat ttttccacca ttgtgaatac
     2281 cacaattcga tgctatcgta tcgacaaagc actttaccca gagaaattcg aagcctccaa
     2341 cccgcactta ccgccacata tggcgttatc aatgagcctc acggcagggc ttttttccaa
     2401 aacattggtg tttttttgtt ctctgatcta ctttaagttg acgcttttcg aaaatggact
     2461 atgtctatcc aacgatatgg aagttggatg gtcggatttg acgaaattga ctgttccttt
     2521 agacaaggat ctatctttag gtacagcaat gagcttgtac tcgtcaatct tcgatcgctt
     2581 gttcacggtc ggaaataaag aattaatacg cacaatgcat agatcatcac aatttgtgat
     2641 agagctagct attgaaagga cctataggac tattcttggc aacgttattg aattcagaaa
     2701 gttgaccgag gaaacatggc tagcacaaat caaacaagaa ctagatccgc agagcgataa
     2761 tccatcatca gaagctaaaa ttgtgagtga taggcagcgg gatctcagcc ttgcagttcc
     2821 aacgcccacg ccctcaataa ttccaatgtt accctcgcct ggtgaaacta agaaccatgc
     2881 aaagagtcaa tcagagatca ttcagatgct gacagatgaa ttttgggcaa attataactc
     2941 aggctgggaa gagttgatta atcaatctga attttccact cttttcgatg attacaaaga
     3001 caataaccca gctttcttgt acaaagtggt tgatagagtg tcgcatagtg atactaattc
     3061 tagcaaaccc gaaaacacga aggagaacct ttcaaaaagc tcttggaggc aggaatggct
     3121 tgcaaatttg aaacttattt ccgtttcgtt ggttgatgag ttcccttcgg agctttccga
     3181 cagtgataga caagtaatta acgaaaaaat gcagttactt aaagatatat ttgctaacaa
     3241 ccttaaatca gcaatttcca ataattttag agagagtgac atcattatac tgaaaggtga
     3301 aatagaagat tacccaatga gttctgaaat taagatttac tacaacgaat tacagaacaa
     3361 gcctgatgca aaaaaagcca gattttggtc atttatgaag actcagagat ttgtttccaa
     3421 catgggattt gatattcgga agtcctgtga acctgtttct atatccacgt cggtcaaacc
     3481 acatgtggtc gagcccgagc atatggcaga tgccaaaatt atgcctaagg atgtactgca
     3541 aattacaaaa aaaccgttaa tggttaaaaa tgtgaagcct tcttctccgc cagatgtgaa
     3601 gtctttggtg cagctgagca caatggaaac gaaaacccta ccagaaaaga agcaattcga
     3661 cagtattttt aattctaaca aagcaaaaat aatccctgga aatgggaagc acgcatcaga
     3721 aaacatctca ctctctttct caagacctgc gtcctacggc tatttttctg ttggaaaaag
     3781 ggttcgggga tcctctagag gtgaacaaaa gttgatttct gaagaagatt tgaacggtga
     3841 acaaaagcta atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga
     3901 cttgaacgga tcctctagag tcgactgaat cgtagatact gaaaaacccc gcaagttcac
     3961 ttcaactgtg catcgtgcac catctcaatt tctttcattt atacatcgtt ttgccttctt
     4021 ttatgtaact atactcctct aagtttcaat cttggccatg taacctctga tctatagaat
     4081 tttttaaatg actagaatta atgcccatct tttttttgga cctaaattct tcatgaaaat
     4141 atattacgag ggcttattca gaagctttgg acttcttcgc cagaggtttg gtcaagtctc
     4201 caatcaaggt tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc
     4261 aaatcgttgg tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga
     4321 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     4381 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     4441 ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg ccgagcaaat
     4501 gcctgcaaat cgctccccat ttcacccaat tgtagatatg ctaactccag caatgagttg
     4561 atgaatctcg gtgtgtattt tatgtcctca gaggacaaca cctgttgtaa tcgttcttcc
     4621 acacggatcc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4681 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4741 ttcacaccgc ataatcgctg ggccattctc atgaagaata tcttgaattt attgtcatat
     4801 tactagttgg tgtggaagtc ctaatatcgg tgatcaatat agtggttgac atgctggcta
     4861 gtcaacattg agccttttga tcatgcaaat atattacggt attttacaat caaatatcaa
     4921 acttaactat tgactttata acttatttag gtggtaacat tcttataaaa aagaaaaaaa
     4981 ttactgcaaa acagtactag cttttaactt gtatcctagg ttatctatgc tgtctcacca
     5041 tagagaatat tacctatttc agaatgtatg tccatgattc gccgggtaaa tacatataat
     5101 acacaaatct ggcttaataa agtctataat atatctcata aagaagtgct aaattggcta
     5161 gtgctatata tttttaagaa aatttctttt gactaagtcc atatcgactt tgtaaaagtt
     5221 cacattagca tacatatatt acacgagcca gaaatagtaa cttttgccta aatcacaaat
     5281 tgcaaaattt aattgcttgc aaaaggtcac atgcttataa tcaacttttt taaaaattta
     5341 aaatactttt ttatttttta tttttaaaca taaatgaaat aatttattta ttgtttatga
     5401 ttaccgaaac ataaaacctg ctcaagaaaa agaaactgtt ttgtccttgg aaaaaaagca
     5461 ctacctagga gcggccaaaa tgccgaggct ttcatagctt aaactcttta cagaaaatag
     5521 gcattataga tcagttcgag ttttcttatt cttccttccg gttttatcgt cacagtttta
     5581 cagtaaataa gtatcacctc ttagagttcg atgataagct gtcaaacatg agaattaatt
     5641 ccacatgtta aaatagtgaa ggagcatgtt cggcacacag tggaccgaac gtggggtaag
     5701 tgcactaggg tccggttaaa cggatctcgc attgatgagg caacgctaat tatcaacata
     5761 tagattgtta tctatctgca tgaacacgaa atctttactt gacgacttga ggctgatggt
     5821 gtttatgcaa agaaaccact gtgtttaata tgtgtcactg tttgatatta ctgtcagcgt
     5881 agaagataat agtaaaagcg gttaataagt gtatttgaga taagtgtgat aaagttttta
     5941 cagcgaaaag acgataaata caagaaaatg attacgagga tacggagaga ggtatgtaca
     6001 tgtgtattta tatactaagc tgccggcggt tgtttgcaag accgagaaaa ggctagcaag
     6061 aatcgggtca ttgtagcgta tgcgcctgtg aacattctct tcaacaagtt tgattccatt
     6121 gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt ttcctgtaca atcaatcaaa
     6181 aagccaaatg atttagcatt atctttacat cttgttattt tacagatttt atgtttagat
     6241 cttttatgct tgcttttcaa aaggcttgca ggcaagtgca caaacaatac ttaaataaat
     6301 actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt
     6361 cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc
     6421 taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagctct
     6481 cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg
     6541 tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac
     6601 tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga
     6661 actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat
     6721 cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt
     6781 tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa
     6841 ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg
     6901 aatctagtgc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac
     6961 cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca
     7021 cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt
     7081 tttcgaccga attaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8161 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8461 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9001 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9061 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9121 accgcctctc cccgcgcgtt ggccgattca ttaatgcagg atccgggatc gaagaaatga
     9181 tggtaaatga aataggaaat caaggagcat gaaggcaaaa gacaaatata agggtcgaac
     9241 gaaaaataaa gtgaaaagtg ttgatatgat gtatttggct ttgcggcgcc gaaaaaacga
     9301 gtttacgcaa ttgcacaatc atgctgactc tgtggcggac ccgcgctctt gccggcccgg
     9361 cgataacgct gggcgtgagg ctgtgcccgg cggagttttt tgcgcctgca ttttccaagg
     9421 tttaccctgc gctaaggggc gagattggag aagcaataag aatgccggtt ggggttgcga
     9481 tgatgacgac cacgacaact ggtgtcatta tttaagttgc cgaaagaacc tgagtgcatt
     9541 tgcaacatga gtatactaga agaatgagcc aagacttgcg agacgcgagt ttgccggtgg
     9601 tgcgaacaat agagcgacca tgaccttgaa ggtgagacgc gcataaccgc tagagtactt
     9661 tgaagaggaa acagcaatag ggttgctacc agtataaata gacaggtaca tacaacactg
     9721 gaaatggttg tctgtttgag tacgctttca attcatttgg gtgtgcactt tattatgtta
     9781 caatatggaa gggaacttta cacttctcct atgcacatat attaattaaa gtccaatgct
     9841 agtagagaag gggggtaaca cccctccgcg ctcttttccg atttttttct aaaccgtgga
     9901 atatttcgga tatccttttg ttgtttccgg gtgtacaata tggacttcct cttttctggc
     9961 aaccaaaccc atacatcggg attcctataa taccttcgtt ggtctcccta acatgtaggt
    10021 ggcggagggg agatatacaa tagaacagat accagacaag acataatggg ctaaacaaga
    10081 ctacaccaat tacactgcct cattgatg
//
