LOCUS       sequence-422167-        6522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Facilitates genomic integration of the reporter with upstream 
            binding sites for ZF-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6522)
  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 6522)
  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_mNeonGreen
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2403..2810
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2811..3905
                     /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(4606..5397)
                     /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      5827..6415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6316..6335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caatacccag aagaacagta aaataaagca aggtacgtga aattaatatt tttaaatggt
       61 tctaaccgat gccgaagaac tgcgcagtcc ggttataacg tctgacatgt ccttttttga
      121 tttggaatcc aaccactcaa gtgactctgt tcatttactt tgcgaaaaat atacccacaa
      181 attgcccatc gaaagtgaat cgcaaaccac cttcagactg gcaccgacaa agcaaagatt
      241 atacagacag agtactttat acgtaccgtt aagtctcaag caaagggttt tcttatttac
      301 tgaacgggta aagagtatct gggccggctt gccaagatgc aaaccgaata agtatttcaa
      361 agttgcattt gccttagccg tcctgacacc attggctatt tggatatttt atattgactt
      421 tcgtgtacat tgatcacatc gactgttcta ttggcaaatg aaccacgggc attgactatt
      481 tttcaggtta ctactatata ttatcatcac gggcaaggat tgtaccctga attcgcatct
      541 agactgatga gacgtggtag agccacaaac agccggtaca agcaacgatc tccaggacca
      601 tctgaatcat gcgcggatga cacgaactca cgacggcgat cacagacatt aacccacagt
      661 acagacactg cgacaacgtg gcaattcgtc gcaatacaac ggaattcaga cgctgctctt
      721 cagacgctgc tctgcagacg ctgctctcca gacgctgctc ggatcccaga tccgccaggc
      781 gtgtatatat agcgtggatg gccaggcaac tttagtgctg acacatacag gcatatatat
      841 atgtgtgcga cgacacatga tcatatggca tgcatgtgct ctgtatgtat ataaaactct
      901 tgttttcttc ttttctctaa atattctttc cttatacatt aggacctttg cagcataaat
      961 tactatactt ctatagacac acaaacacaa atacacacac taatctagat attaaatatg
     1021 gtttcaaagg gtgaagagga taacatggca agtttgccgg ccacacatga acttcacatt
     1081 ttcgggagca taaatggcgt tgattttgac atggtcggtc aggggactgg gaaccccaac
     1141 gacggatacg aagaactcaa cttgaaatct accaaaggtg atctccaatt ttcaccgtgg
     1201 attcttgttc cgcacattgg gtatggattt catcagtatt tgccataccc tgatggcatg
     1261 tcacccttcc aggccgctat ggtcgatggt agcggctacc aggtacaccg cacaatgcag
     1321 tttgaagacg gtgcatcact gaccgtgaac taccgatata cctatgaggg ttcccatatt
     1381 aaaggagaag ctcaagtaaa gggaacggga ttcccggccg atggtcccgt gatgactaac
     1441 tccttgacag cagcagactg gtgtcgctct aagaagactt atcccaacga taaaaccatt
     1501 ataagtacat ttaaatggag ttatacaacg ggcaacggta agaggtacag aagcacggca
     1561 cggaccactt acaccttcgc taagccgatg gccgcaaact acttgaagaa ccaaccgatg
     1621 tacgtattcc gaaagacgga actgaaacat agcaaaactg aactgaattt caaggagtgg
     1681 cagaaagcgt ttactgatgt catgggtatg gacgagctgt ataagggatc ctaactcgag
     1741 ataaagcaat cttgatgagg ataatgattt ttttttgaat atacataaat actaccgttt
     1801 ttctgctaga ttttgtgatg acgtaaataa gtacatatta ctttttaagc caagacaaga
     1861 ttaagcatta actttaccct tttctttcta agtttcaata ttagttatca ctgtttaaaa
     1921 gttatggcga gaacgtcggc ggttaaaata tattaccctg aacggctgga aatctgctcg
     1981 tcagtggtgc tcacactgac gaatcatgta cagatcatac cgatgactgc ctggcgactc
     2041 acaactaagc aagacagccg gaaccagcgc cggcgaacac cactgcatat atggcatatc
     2101 acaacagtcc acgtctcaag cagttacaga gatgttacga accactagtg cactgcagta
     2161 catcgaggag aacttctagt atatctacat acctaatatt attgccttat taaaaatgga
     2221 atcccaacaa ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt
     2281 gttcatgtgt gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag
     2341 taattggttg tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag
     2401 ttaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat
     2461 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
     2521 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
     2581 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
     2641 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
     2701 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct
     2761 tttacatttc agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc
     2821 ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca
     2881 ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc
     2941 atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg
     3001 aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggaa
     3061 ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg
     3121 ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc
     3181 cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact
     3241 ttggtaagag aaaggaggac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg
     3301 ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac
     3361 cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa
     3421 aaactgtgga ggaaaccatc aagaacgaat ttcctacatt gaaggttcaa catcaattga
     3481 ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca
     3541 ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg
     3601 gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt
     3661 acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca
     3721 ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg
     3781 ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg
     3841 gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg
     3901 cttaacctgg aggacccttc tctttagact attctactct tatgcacgta aaaaattcta
     3961 ggaaatatgt attaactagg agtaaaataa ccggctagtg gcattcatat agccgtctgt
     4021 ttacatctac atcacacatt tcgagtgtat atctcgcaac gttggcgtta aataggcagg
     4081 agtactcgta tcgcatgtcg gtgcgacacg aaattacaaa atggaatatg ttcatagggt
     4141 agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta
     4201 aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc
     4261 actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa
     4321 aatcatgaaa ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa
     4381 tacaacaaat aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt
     4441 cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg
     4501 ccttaacgac gtagtcgacc tcctcttcag tactaaatct accaatacca aatctgatgg
     4561 aagaatgggc taatgcatca tccttaccca gcgccgagcg gccgcttatt tgccgactac
     4621 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     4681 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     4741 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     4801 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     4861 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     4921 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     4981 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     5041 cgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     5101 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     5161 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     5221 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     5281 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     5341 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     5401 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     5461 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     5521 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     5581 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     5641 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     5701 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg tcatgaccaa
     5761 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6481 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gc
//
