LOCUS       sequence-422184-        6662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Facilitates genomic integration of the reporter for screen 1: ZF-1 
            followed by ZF-2 upstream of the reporter gene. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6662)
  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 6662)
  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_ZF2_ZF1_mNeonGreen
FEATURES             Location/Qualifiers
     source          1..6662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2019..2426
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2427..3521
                     /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(4222..5013)
                     /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      5443..6031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5932..5951
                     /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 gacgaacggc attcattatg ggagattgat tatgggagat ggattatggg
      241 agatcgatta tgggagagct agcagacgct gctcttcaga cgctgctctg cagacgctgc
      301 tctccagacg ctgctcgaat tcagagtgag gacttgagag tgaggactgg agagtgagga
      361 ctcgagagtg aggacggatc ccagatccgc caggcgtgta tatatagcgt ggatggccag
      421 gcaactttag tgctgacaca tacaggcata tatatatgtg tgcgacgaca catgatcata
      481 tggcatgcat gtgctctgta tgtatataaa actcttgttt tcttcttttc tctaaatatt
      541 ctttccttat acattaggac ctttgcagca taaattacta tacttctata gacacacaaa
      601 cacaaataca cacactaatc tagatattaa atatggtttc aaagggtgaa gaggataaca
      661 tggcaagttt gccggccaca catgaacttc acattttcgg gagcataaat ggcgttgatt
      721 ttgacatggt cggtcagggg actgggaacc ccaacgacgg atacgaagaa ctcaacttga
      781 aatctaccaa aggtgatctc caattttcac cgtggattct tgttccgcac attgggtatg
      841 gatttcatca gtatttgcca taccctgatg gcatgtcacc cttccaggcc gctatggtcg
      901 atggtagcgg ctaccaggta caccgcacaa tgcagtttga agacggtgca tcactgaccg
      961 tgaactaccg atatacctat gagggttccc atattaaagg agaagctcaa gtaaagggaa
     1021 cgggattccc ggccgatggt cccgtgatga ctaactcctt gacagcagca gactggtgtc
     1081 gctctaagaa gacttatccc aacgataaaa ccattataag tacatttaaa tggagttata
     1141 caacgggcaa cggtaagagg tacagaagca cggcacggac cacttacacc ttcgctaagc
     1201 cgatggccgc aaactacttg aagaaccaac cgatgtacgt attccgaaag acggaactga
     1261 aacatagcaa aactgaactg aatttcaagg agtggcagaa agcgtttact gatgtcatgg
     1321 gtatggacga gctgtataag ggatcctaac tcgaggccaa ttggtgcggc aattgataat
     1381 aacgaaaatg tcttttaatg atctgggtat aatgaggaat tttccgaacg tttttacttt
     1441 atatatatat atacatgtaa catatattct atacgctata gagaaaggaa atttttcaat
     1501 taaaaaaaaa atagagaaag agtttcactt cttgattatc gctaacacta atggttgaag
     1561 tactgctact ttaattttat gctggaaatc tgctcgtcag tggtgctcac actgacgaat
     1621 catgtacaga tcataccgat gactgcctgg cgactcacaa ctaagcaaga cagccggaac
     1681 cagcgccggc gaacaccact gcatatatgg catatcacaa cagtccacgt ctcaagcagt
     1741 tacagagatg ttacgaacca ctagtgcact gcagtacatc gaggagaact tctagtatat
     1801 ctacatacct aatattattg ccttattaaa aatggaatcc caacaattac atcaaaatcc
     1861 acattctctt caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta
     1921 tatttatagg ataattatac tctatttctc aacaagtaat tggttgtttg gccgagcggt
     1981 ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa ctgtgggaat actcaggtat
     2041 cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca acataacgag
     2101 aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt
     2161 tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg
     2221 tgagagcgcc ggaaccggct tttcatatag aatagagaag cgttcatgac taaatgcttg
     2281 catcacaata cttgaagttg acaatattat ttaaggacct attgtttttt ccaataggtg
     2341 gttagcaatc gtcttacttt ctaacttttc ttacctttta catttcagca atatatatat
     2401 atatatttca aggatatacc attctaatgt ctgcccctaa gaagatcgtc gttttgccag
     2461 gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg
     2521 atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg
     2581 atgctacagg tgttccactt ccagatgagg cgctggaagc ctccaagaag gctgatgccg
     2641 ttttgttagg tgctgtgggt ggtcctaaat ggggaaccgg tagtgttaga cctgaacaag
     2701 gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg
     2761 catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact
     2821 tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag gaggacgatg
     2881 gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa
     2941 gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata
     3001 aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga
     3061 acgaatttcc tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag
     3121 ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca
     3181 tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg
     3241 cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc
     3301 cagatttgcc aaagaataag gtcaacccta tcgccactat cttgtctgct gcaatgatgt
     3361 tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg
     3421 ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg
     3481 gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta acctggagga cccttctctt
     3541 tagactattc tactcttatg cacgtaaaaa attctaggaa atatgtatta actaggagta
     3601 aaataaccgg ctagtggcat tcatatagcc gtctgtttac atctacatca cacatttcga
     3661 gtgtatatct cgcaacgttg gcgttaaata ggcaggagta ctcgtatcgc atgtcggtgc
     3721 gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta
     3781 catacattta tcaagaagga gaaaaaggag gatgtaaagg aatacaggta agcaaattga
     3841 tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaat attgacaagg
     3901 aggagggcac cacacaaaaa gttaggtgta acagaaaatc atgaaactat gattcctaat
     3961 ttatatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat
     4021 gacctgacca tttgatggag tttaagtcaa taccttcttg aaccatttcc cataatggtg
     4081 aaagttccct caagaatttt actctgtcag aaacggcctt aacgacgtag tcgacctcct
     4141 cttcagtact aaatctacca ataccaaatc tgatggaaga atgggctaat gcatcatcct
     4201 tacccagcgc cgagcggccg cttatttgcc gactaccttg gtgatctcgc ctttcacgta
     4261 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     4321 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     4381 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     4441 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     4501 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     4561 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     4621 cagatcaatg tcgatcgtgg ctggctcgaa gatacccgca agaatgtcat tgcgctgcca
     4681 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     4741 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     4801 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     4861 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     4921 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     4981 agtcgatact tcggcgatca ccgcttccct cataatgttt aactttgttt tagggcgact
     5041 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     5101 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
     5161 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     5221 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     5281 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gtagcagcga
     5341 agtcgaggca tttctgtcct ggctggtcat gaccaaaatc ccttaacgtg agttttcgtt
     5401 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5461 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5521 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5581 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5641 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5701 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5761 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5821 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5881 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5941 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6001 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6061 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6121 ggataaccgt gcggccgcca atacccagaa gaacagtaaa ataaagcaag gtacgtgaaa
     6181 ttaatatttt taaatggttc taaccgatgc cgaagaactg cgcagtccgg ttataacgtc
     6241 tgacatgtcc ttttttgatt tggaatccaa ccactcaagt gactctgttc atttactttg
     6301 cgaaaaatat acccacaaat tgcccatcga aagtgaatcg caaaccacct tcagactggc
     6361 accgacaaag caaagattat acagacagag tactttatac gtaccgttaa gtctcaagca
     6421 aagggttttc ttatttactg aacgggtaaa gagtatctgg gccggcttgc caagatgcaa
     6481 accgaataag tatttcaaag ttgcatttgc cttagccgtc ctgacaccat tggctatttg
     6541 gatattttat attgactttc gtgtacattg atcacatcga ctgttctatt ggcaaatgaa
     6601 ccacgggcat tgactatttt tcaggttact actatatatt atcatcacgg gcaaggattg
     6661 ta
//
