LOCUS       sequence-422166-        6615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Facilitates genomic integration of the reporter for screen 2: ZF-1 
            upstream of the reporter gene and ZF-2 downstream. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  AUTHORS   Alcantar MA, English MA, Valeri JA, Collins JJ
  TITLE     A high-throughput synthetic biology approach for studying 
            combinatorial chromatin-based transcriptional regulation.
  JOURNAL   Mol Cell. 2024 Jun 20;84(12):2382-2396.e9. doi: 
            10.1016/j.molcel.2024.05.025.
  PUBMED    38906116
REFERENCE   2  (bases 1 to 6615)
  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: 
            "10.1016/j.molcel.2024.05"; journalName: "Mol Cell"; date: 
            "2024-06-20- 20"; volume: "84"; issue: "12"; pages: "2382-2396.e9"
COMMENT     Sequence Label: pReporter_ZF1_mNeonGreen_ZF2
FEATURES             Location/Qualifiers
     source          1..6615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1972..2379
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2380..3474
                     /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"
     CDS             complement(4175..4966)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      5396..5984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5885..5904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgga attcagagtg aggacttgag agtgaggact ggagagtgag
      241 gactcgagag tgaggacgga tcccagatcc gccaggcgtg tatatatagc gtggatggcc
      301 aggcaacttt agtgctgaca catacaggca tatatatatg tgtgcgacga cacatgatca
      361 tatggcatgc atgtgctctg tatgtatata aaactcttgt tttcttcttt tctctaaata
      421 ttctttcctt atacattagg acctttgcag cataaattac tatacttcta tagacacaca
      481 aacacaaata cacacactaa tctagatatt aaatatggtt tcaaagggtg aagaggataa
      541 catggcaagt ttgccggcca cacatgaact tcacattttc gggagcataa atggcgttga
      601 ttttgacatg gtcggtcagg ggactgggaa ccccaacgac ggatacgaag aactcaactt
      661 gaaatctacc aaaggtgatc tccaattttc accgtggatt cttgttccgc acattgggta
      721 tggatttcat cagtatttgc cataccctga tggcatgtca cccttccagg ccgctatggt
      781 cgatggtagc ggctaccagg tacaccgcac aatgcagttt gaagacggtg catcactgac
      841 cgtgaactac cgatatacct atgagggttc ccatattaaa ggagaagctc aagtaaaggg
      901 aacgggattc ccggccgatg gtcccgtgat gactaactcc ttgacagcag cagactggtg
      961 tcgctctaag aagacttatc ccaacgataa aaccattata agtacattta aatggagtta
     1021 tacaacgggc aacggtaaga ggtacagaag cacggcacgg accacttaca ccttcgctaa
     1081 gccgatggcc gcaaactact tgaagaacca accgatgtac gtattccgaa agacggaact
     1141 gaaacatagc aaaactgaac tgaatttcaa ggagtggcag aaagcgttta ctgatgtcat
     1201 gggtatggac gagctgtata agggatccta actcgaggcc aattggtgcg gcaattgata
     1261 ataacgaaaa tgtcttttaa tgatctgggt ataatgagga attttccgaa cgtttttact
     1321 ttatatatat atatacatgt aacatatatt ctatacgcta tagagaaagg aaatttttca
     1381 attaaaaaaa aaatagagaa agagtttcac ttcttgatta tcgctaacac taatggttga
     1441 agtactgcta ctttaatttt ataacggcta gcagacgctg ctcttcagac gctgctctgc
     1501 agacgctgct ctccagacgc tgctcggatc cgggctggaa atctgctcgt cagtggtgct
     1561 cacactgacg aatcatgtac agatcatacc gatgactgcc tggcgactca caactaagca
     1621 agacagccgg aaccagcgcc ggcgaacacc actgcatata tggcatatca caacagtcca
     1681 cgtctcaagc agttacagag atgttacgaa ccactagtgc actgcagtac atcgaggaga
     1741 acttctagta tatctacata cctaatatta ttgccttatt aaaaatggaa tcccaacaat
     1801 tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg
     1861 ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt aattggttgt
     1921 ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt taactgtggg
     1981 aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt atttttttcc
     2041 tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga tgactggaaa
     2101 ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga aaaattggga
     2161 gaaaaaggaa aggtgagagc gccggaaccg gcttttcata tagaatagag aagcgttcat
     2221 gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga cctattgttt
     2281 tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt ttacatttca
     2341 gcaatatata tatatatatt tcaaggatat accattctaa tgtctgcccc taagaagatc
     2401 gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt
     2461 aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt
     2521 ggtgctgcta tcgatgctac aggtgttcca cttccagatg aggcgctgga agcctccaag
     2581 aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggaac cggtagtgtt
     2641 agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga
     2701 ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct
     2761 aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga
     2821 aaggaggacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg
     2881 caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt
     2941 tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag
     3001 gaaaccatca agaacgaatt tcctacattg aaggttcaac atcaattgat tgattctgcc
     3061 gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg
     3121 tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca
     3181 tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc
     3241 cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac tatcttgtct
     3301 gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat
     3361 gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt
     3421 accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaacctgga
     3481 ggacccttct ctttagacta ttctactctt atgcacgtaa aaaattctag gaaatatgta
     3541 ttaactagga gtaaaataac cggctagtgg cattcatata gccgtctgtt tacatctaca
     3601 tcacacattt cgagtgtata tctcgcaacg ttggcgttaa ataggcagga gtactcgtat
     3661 cgcatgtcgg tgcgacacga aattacaaaa tggaatatgt tcatagggta gacgaaacta
     3721 tatacgcaat ctacatacat ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag
     3781 gtaagcaaat tgatactaat ggctcaacgt gataaggaaa aagaattgca ctttaacatt
     3841 aatattgaca aggaggaggg caccacacaa aaagttaggt gtaacagaaa atcatgaaac
     3901 tatgattcct aatttatata ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata
     3961 aaaaacactc aatgacctga ccatttgatg gagtttaagt caataccttc ttgaaccatt
     4021 tcccataatg gtgaaagttc cctcaagaat tttactctgt cagaaacggc cttaacgacg
     4081 tagtcgacct cctcttcagt actaaatcta ccaataccaa atctgatgga agaatgggct
     4141 aatgcatcat ccttacccag cgccgagcgg ccgcttattt gccgactacc ttggtgatct
     4201 cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
     4261 cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca
     4321 ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc
     4381 tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg
     4441 gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg
     4501 gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg
     4561 tcagcaagat agccagatca atgtcgatcg tggctggctc gaagataccc gcaagaatgt
     4621 cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga
     4681 tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg
     4741 aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta
     4801 cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg
     4861 agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta
     4921 cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg
     4981 ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
     5041 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac
     5101 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     5161 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg
     5221 cttatgtcca ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     5281 tgggtagcag cgaagtcgag gcatttctgt cctggctggt catgaccaaa atcccttaac
     5341 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5401 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5461 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5521 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5581 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5641 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5701 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5761 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5821 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5881 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5941 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6001 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6061 cccctgattc tgtggataac cgtgcggccg ccaataccca gaagaacagt aaaataaagc
     6121 aaggtacgtg aaattaatat ttttaaatgg ttctaaccga tgccgaagaa ctgcgcagtc
     6181 cggttataac gtctgacatg tccttttttg atttggaatc caaccactca agtgactctg
     6241 ttcatttact ttgcgaaaaa tatacccaca aattgcccat cgaaagtgaa tcgcaaacca
     6301 ccttcagact ggcaccgaca aagcaaagat tatacagaca gagtacttta tacgtaccgt
     6361 taagtctcaa gcaaagggtt ttcttattta ctgaacgggt aaagagtatc tgggccggct
     6421 tgccaagatg caaaccgaat aagtatttca aagttgcatt tgccttagcc gtcctgacac
     6481 cattggctat ttggatattt tatattgact ttcgtgtaca ttgatcacat cgactgttct
     6541 attggcaaat gaaccacggg cattgactat ttttcaggtt actactatat attatcatca
     6601 cgggcaagga ttgta
//
