LOCUS       sequence-434484-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene (4 operators) for zinc finger activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  AUTHORS   Bragdon MDJ, Patel N, Chuang J, Levien E, Bashor CJ, Khalil AS
  TITLE     Cooperative assembly confers regulatory specificity and long-term 
            genetic circuit stability.
  JOURNAL   Cell. 2023 Aug 31;186(18):3810-3825.e18. doi: 
            10.1016/j.cell.2023.07.012. Epub 2023 Aug 7.
  PUBMED    37552983
REFERENCE   2  (bases 1 to 5579)
  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.cell.2023.07.012"; journalName: "Cell"; date: 
            "2023-08-31- 31"; volume: "186"; issue: "18"; pages: "3810-3825.e18"
COMMENT     Sequence Label: pKL1-100
FEATURES             Location/Qualifiers
     source          1..5579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             313..1026
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEGGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1042..1231
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             1586..2389
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(join(3098..3889,3890..3958))
                     /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     3721..3740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3959..4063)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggaattc agacgctgct ctcgagacgc tgctctcgag acgctgctct cgagacgctg
       61 ctcggatccc agatccgcca ggcgtgtata tatagcgtgg atggccaggc aactttagtg
      121 ctgacacata caggcatata tatatgtgtg cgacgacaca tgatcatatg gcatgcatgt
      181 gctctgtatg tatataaaac tcttgttttc ttcttttctc taaatattct ttccttatac
      241 attaggacct ttgcagcata aattactata cttctataga cacacaaaca caaatacaca
      301 cactaaagat ctatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
      361 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
      421 gctacttacg gtaaattgac cttaaaattg atttgtacta ctggtaaatt gccagttcca
      481 tggccaacct tagtcactac tttaggttat ggtttgcaat gttttgctag atacccagat
      541 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
      601 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
      661 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaagg tggtaacatt
      721 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac tgctgacaaa
      781 caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga tggtggtgtt
      841 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
      901 gacaaccatt acttatccta tcaatctgcc ttatccaaag atccaaacga aaagagagat
      961 cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat ggatgaattg
     1021 tacaaaggat cctaactcga gatccgctct aaccgaaaag gaaggagtta gacaacctga
     1081 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     1141 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     1201 cttgcttgag aaggttttgg gacgctcgaa ggctgccaat gagacgacgg ggtcatcacg
     1261 gctcatcatg cgccaaacaa atgtgtgcaa tacacgctcg gatgactgca tgatgaccgc
     1321 actgactggg gacagcagat ccacctaagc ctgtgagaga agcagacacc cgacagatca
     1381 aggcagttaa ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat
     1441 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
     1501 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
     1561 aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
     1621 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
     1681 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
     1741 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
     1801 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag
     1861 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
     1921 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
     1981 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
     2041 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
     2101 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
     2161 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc
     2221 attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat
     2281 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     2341 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     2401 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     2461 attacccgag tagagcactt gaatccactg ccccgggaat ctcggtcgta atgatttcta
     2521 taatgacgaa aaaaaaaaaa ttggaaagaa aaagcttcat ggcctttata aaaaggaact
     2581 atccaatacc tcgccagaac caagtaacag tattttacgg ggcacaaatc aagaacaata
     2641 agacaggact gtaaagatgg acgcattgaa ctccaaagaa caacaagagt tccaaaaagt
     2701 agtggaacaa aagcaaatga aggatttcat gcgtttgtac tctaatctgg tagaaagatg
     2761 tttcacagac tgtgtcaatg acttcacaac atcaaagcta accaataagg aacaaacatg
     2821 catcatgaag tgctcagaaa agttcttgaa gcatagcgaa cgtgtagggc agcgtttcca
     2881 agaacaaaac gctgccttgg gacaaggctt gggccgataa ggtgtactgg cgtatatata
     2941 tctaattatg tatctctggt gtagcccatt tttagcatgt aaatataaag accgagcggc
     3001 cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg
     3241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4081 tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtgcggccg
     4861 ccaatgctaa attcgagtga aacacaggaa gatcagaaaa tcctcatttc atccatatta
     4921 acaataattt caaatgttta tttgcattat ttgaaactag gcaagacaag caacgaaacg
     4981 tttttgaaaa ttttgagtat tttcaataaa tttgtagagg actcagatat tgaaaaaaag
     5041 ctacagcaat taatacttga taagaagagt attgagaagg gcaacggttc atcatctcat
     5101 ggatctgcac atgaacaaac accagagtca aacgacgttg aaattgaggc tactgcgcca
     5161 attgatgaca atacagacga tgataacaaa ccgaagttat ctgatgtaga aaaggattaa
     5221 agatgctaag agatagtgat gatatttcat aaataatgta attctatata tgttaattac
     5281 cttttttgcg aggcatattt atggtgaagg ataagttttg accatcaaag aaggttaatg
     5341 tggctgtggt ttcagggtcc ataaagccca catggataac attacccctg aattcgcatc
     5401 tagatggtag agccacaaac agccggtaca agcaacgatc tccaggacca tctgaatcat
     5461 gcgcggatga cacgaactca cgacggcgat cacagacatt aacccacagt acagacactg
     5521 cgacaacgtg gcaattcgtc gcaataccgt ctcactgaac tggccgataa ttgcagacg
//
