LOCUS       sequence-381350-       10048 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 10048)
  TITLE     A Novel Mode of Half Site Binding in Budding Yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10048)
  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            "Unpublished"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_LYS14_SIR4
FEATURES             Location/Qualifiers
     source          1..10048
                     /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(2946..2970)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3743..3772
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3779..3808
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3815..3844
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      4239..4426
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4294..4318)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    5256..6418
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      5581..6418
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(6257..6931)
                     /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(6932..7033)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7053..7071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7139..7243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7244..7312,7313..8104)
                     /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(7462..7481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8275..8863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8764..8783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9017..9034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(9852..10048,1..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 caaaatgttc gaatctgtca atttggatga
      601 aaattctcca gaggatagag aactggctaa ggttttgtcc ccaccgggta gttatctttc
      661 tccagcttca ctagacagtg gttcatcgtt cacaaatagt gggacttcaa ctagttgctt
      721 tgaaccaaaa aacaatctac cctccttatc gtttttaaat gctagagcgg gatctttagg
      781 tggcattttt aatcataaac agatgacctc gccgtcaaat tcgaatatag gaggcgaaaa
      841 tgtagaatcg accacaagta gtaatgacgg cagtaatgaa aatgcaggtc atcccactac
      901 ttcagagcaa gatcaaaatg cagatcaccc cactatttca caggcagatg ataatggaca
      961 ctcatcgttg acccctaatc ctgctgtaac ttcaactgtt acggataaga agggaaatac
     1021 agtaaaacgg aagtactcca gaaatgggtg ctccgagtgt aagagacgaa gaatgaaatg
     1081 tgatgaaact aaaccgactt gttggcagtg cgcaagattg aatcgtcagt gcgtttacgt
     1141 tttaaaccca aaaaataaga aaagaagaac ctcaaatgct caaagagtaa aagaatttag
     1201 gaagcacagt actagcctcg ataatgatca taataatgct cgcaagcgtc agcattcgtc
     1261 atgcaaggct gagaagaaga aaaaagtacg acaaaattta agtgaagaca ccacggaccc
     1321 taaaccgatt acagataacg gcaagaatgt acctttggat gaaatagaat ccctagagat
     1381 tccaaatttg gaccttacca ccacaatgaa tggatatgac gttaatttac taatgcagaa
     1441 cttgaatgat atggttaata tgaaacttca tgactcttac ttattgaatg aagaactgaa
     1501 aggattagat ttgcccgatt tggacatacc tgaacttttg cccgcttcga atgtgaacag
     1561 tagcgtacca atatcctttc tggtgaataa tgtaataact ttcaacacaa aactaagctc
     1621 gttcaaacta ggtggaatcc acgataagta tctaaagata ttttactacg attgcttaga
     1681 ttctattgcg ccctttttcc aaaatcaagg gaacccgtta agagatatct tactttcatt
     1741 tgcgaaaaac gaggcctatc ttttatcttc catattagct acgggagcat ctatagcata
     1801 tagaaagtct aataacttgg aagatgaacg aaactattgt gcttatttgt cacattgcct
     1861 gagcctgttg ggtgaacaat tcaagaatga gtcaaacgtt ttaaatagga tagagcctat
     1921 catcctgatg gtcatcatgc ttgcatggga ttgtatttat tcaatgaatt ctcaatggag
     1981 gtctcactta aaaggtgtaa ctgacctttt taaaaaaatc aatgccggga attcctccaa
     2041 agtacttaac gttgcaaaat gttggtttaa agttatggaa acattcgcca gtataagtac
     2101 ggtatttggc ggatctttaa ttgataataa tgaccttgat gcaatttttg atccatatga
     2161 ttaccaatat gttgactcgt taaaattttt aaatattatg actccactaa atgagtttaa
     2221 tcttttaaga ggacataaag aagatttcga tcttgttatt aaagaagttt tcaaatcact
     2281 caatacaatt aggtcaactg aaaaaaatta cttttctaaa gaagaaggtt tgttcactaa
     2341 gaaattagac tatctgcttt tatcatccca aacttcgtcc gaaaagtcca aagaccagat
     2401 atcatatttc aatacgcaga aaattctagt ggaaattgac aagcaattag attacgaatt
     2461 tattgacaaa tctggaatta taccctcaga caatcagtca caccctagaa ttagtaatat
     2521 tcatgataat gccatcgata tggtgacatt aaagaacggt gaagaagttg ccatcagctg
     2581 gtatgatatt tctcatcaga cacgagtttt atctttctta ttaattgtat tattaaaact
     2641 tttaggtatg ccaaaagagt caagtaccat tcaacaagtg gtcaagaaaa tcatgtcttt
     2701 cttcaaattc ttagacagtg actcgccgcc acaaaattca aggacttgtt atagtaattt
     2761 tgctgtattg attgcaggtt tgaatgcaat ggacgaggaa acaagagcga ttgttaaaag
     2821 atattataaa atcaatggtg gaaaatttca aaagctcaca gaacataatc ttaacagact
     2881 ggaaaaggtc tggtatggta aaaatcaaaa ttacaggtta gaagaacaag acgttttaac
     2941 atggaaccca gctttcttgt acaaagtggt tgatagagtg tcgcatagtg atactaattc
     3001 tagcaaaccc gaaaacacga aggagaacct ttcaaaaagc tcttggaggc aggaatggct
     3061 tgcaaatttg aaacttattt ccgtttcgtt ggttgatgag ttcccttcgg agctttccga
     3121 cagtgataga caagtaatta acgaaaaaat gcagttactt aaagatatat ttgctaacaa
     3181 ccttaaatca gcaatttcca ataattttag agagagtgac atcattatac tgaaaggtga
     3241 aatagaagat tacccaatga gttctgaaat taagatttac tacaacgaat tacagaacaa
     3301 gcctgatgca aaaaaagcca gattttggtc atttatgaag actcagagat ttgtttccaa
     3361 catgggattt gatattcgga agtcctgtga acctgtttct atatccacgt cggtcaaacc
     3421 acatgtggtc gagcccgagc atatggcaga tgccaaaatt atgcctaagg atgtactgca
     3481 aattacaaaa aaaccgttaa tggttaaaaa tgtgaagcct tcttctccgc cagatgtgaa
     3541 gtctttggtg cagctgagca caatggaaac gaaaacccta ccagaaaaga agcaattcga
     3601 cagtattttt aattctaaca aagcaaaaat aatccctgga aatgggaagc acgcatcaga
     3661 aaacatctca ctctctttct caagacctgc gtcctacggc tatttttctg ttggaaaaag
     3721 ggttcgggga tcctctagag gtgaacaaaa gttgatttct gaagaagatt tgaacggtga
     3781 acaaaagcta atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga
     3841 cttgaacgga tcctctagag tcgactgaat cgtagatact gaaaaacccc gcaagttcac
     3901 ttcaactgtg catcgtgcac catctcaatt tctttcattt atacatcgtt ttgccttctt
     3961 ttatgtaact atactcctct aagtttcaat cttggccatg taacctctga tctatagaat
     4021 tttttaaatg actagaatta atgcccatct tttttttgga cctaaattct tcatgaaaat
     4081 atattacgag ggcttattca gaagctttgg acttcttcgc cagaggtttg gtcaagtctc
     4141 caatcaaggt tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc
     4201 aaatcgttgg tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga
     4261 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     4321 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     4381 ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg ccgagcaaat
     4441 gcctgcaaat cgctccccat ttcacccaat tgtagatatg ctaactccag caatgagttg
     4501 atgaatctcg gtgtgtattt tatgtcctca gaggacaaca cctgttgtaa tcgttcttcc
     4561 acacggatcc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4621 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4681 ttcacaccgc ataatcgctg ggccattctc atgaagaata tcttgaattt attgtcatat
     4741 tactagttgg tgtggaagtc ctaatatcgg tgatcaatat agtggttgac atgctggcta
     4801 gtcaacattg agccttttga tcatgcaaat atattacggt attttacaat caaatatcaa
     4861 acttaactat tgactttata acttatttag gtggtaacat tcttataaaa aagaaaaaaa
     4921 ttactgcaaa acagtactag cttttaactt gtatcctagg ttatctatgc tgtctcacca
     4981 tagagaatat tacctatttc agaatgtatg tccatgattc gccgggtaaa tacatataat
     5041 acacaaatct ggcttaataa agtctataat atatctcata aagaagtgct aaattggcta
     5101 gtgctatata tttttaagaa aatttctttt gactaagtcc atatcgactt tgtaaaagtt
     5161 cacattagca tacatatatt acacgagcca gaaatagtaa cttttgccta aatcacaaat
     5221 tgcaaaattt aattgcttgc aaaaggtcac atgcttataa tcaacttttt taaaaattta
     5281 aaatactttt ttatttttta tttttaaaca taaatgaaat aatttattta ttgtttatga
     5341 ttaccgaaac ataaaacctg ctcaagaaaa agaaactgtt ttgtccttgg aaaaaaagca
     5401 ctacctagga gcggccaaaa tgccgaggct ttcatagctt aaactcttta cagaaaatag
     5461 gcattataga tcagttcgag ttttcttatt cttccttccg gttttatcgt cacagtttta
     5521 cagtaaataa gtatcacctc ttagagttcg atgataagct gtcaaacatg agaattaatt
     5581 ccacatgtta aaatagtgaa ggagcatgtt cggcacacag tggaccgaac gtggggtaag
     5641 tgcactaggg tccggttaaa cggatctcgc attgatgagg caacgctaat tatcaacata
     5701 tagattgtta tctatctgca tgaacacgaa atctttactt gacgacttga ggctgatggt
     5761 gtttatgcaa agaaaccact gtgtttaata tgtgtcactg tttgatatta ctgtcagcgt
     5821 agaagataat agtaaaagcg gttaataagt gtatttgaga taagtgtgat aaagttttta
     5881 cagcgaaaag acgataaata caagaaaatg attacgagga tacggagaga ggtatgtaca
     5941 tgtgtattta tatactaagc tgccggcggt tgtttgcaag accgagaaaa ggctagcaag
     6001 aatcgggtca ttgtagcgta tgcgcctgtg aacattctct tcaacaagtt tgattccatt
     6061 gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt ttcctgtaca atcaatcaaa
     6121 aagccaaatg atttagcatt atctttacat cttgttattt tacagatttt atgtttagat
     6181 cttttatgct tgcttttcaa aaggcttgca ggcaagtgca caaacaatac ttaaataaat
     6241 actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt
     6301 cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc
     6361 taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagctct
     6421 cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg
     6481 tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac
     6541 tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga
     6601 actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat
     6661 cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt
     6721 tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa
     6781 ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg
     6841 aatctagtgc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac
     6901 cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca
     6961 cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt
     7021 tttcgaccga attaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     7081 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7141 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7201 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7261 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7321 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7381 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7441 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7501 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7561 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7621 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7681 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7741 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7801 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7861 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7921 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7981 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8041 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8101 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8401 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8881 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8941 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9001 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9061 accgcctctc cccgcgcgtt ggccgattca ttaatgcagg atccgggatc gaagaaatga
     9121 tggtaaatga aataggaaat caaggagcat gaaggcaaaa gacaaatata agggtcgaac
     9181 gaaaaataaa gtgaaaagtg ttgatatgat gtatttggct ttgcggcgcc gaaaaaacga
     9241 gtttacgcaa ttgcacaatc atgctgactc tgtggcggac ccgcgctctt gccggcccgg
     9301 cgataacgct gggcgtgagg ctgtgcccgg cggagttttt tgcgcctgca ttttccaagg
     9361 tttaccctgc gctaaggggc gagattggag aagcaataag aatgccggtt ggggttgcga
     9421 tgatgacgac cacgacaact ggtgtcatta tttaagttgc cgaaagaacc tgagtgcatt
     9481 tgcaacatga gtatactaga agaatgagcc aagacttgcg agacgcgagt ttgccggtgg
     9541 tgcgaacaat agagcgacca tgaccttgaa ggtgagacgc gcataaccgc tagagtactt
     9601 tgaagaggaa acagcaatag ggttgctacc agtataaata gacaggtaca tacaacactg
     9661 gaaatggttg tctgtttgag tacgctttca attcatttgg gtgtgcactt tattatgtta
     9721 caatatggaa gggaacttta cacttctcct atgcacatat attaattaaa gtccaatgct
     9781 agtagagaag gggggtaaca cccctccgcg ctcttttccg atttttttct aaaccgtgga
     9841 atatttcgga tatccttttg ttgtttccgg gtgtacaata tggacttcct cttttctggc
     9901 aaccaaaccc atacatcggg attcctataa taccttcgtt ggtctcccta acatgtaggt
     9961 ggcggagggg agatatacaa tagaacagat accagacaag acataatggg ctaaacaaga
    10021 ctacaccaat tacactgcct cattgatg
//
