LOCUS       sequence-434482-        5565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene (3 operator) for zinc finger activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5565)
  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 5565)
  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: pKL1-424
FEATURES             Location/Qualifiers
     source          1..5565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             299..1012
                     /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      1028..1217
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             1572..2375
                     /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(3084..3875,3876..3944))
                     /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     3707..3726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3945..4049)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4152..4740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4641..4660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggaattc agacgctgct ctcgagacgc tgctctcgag acgctgctcg gatcccagat
       61 ccgccaggcg tgtatatata gcgtggatgg ccaggcaact ttagtgctga cacatacagg
      121 catatatata tgtgtgcgac gacacatgat catatggcat gcatgtgctc tgtatgtata
      181 taaaactctt gttttcttct tttctctaaa tattctttcc ttatacatta ggacctttgc
      241 agcataaatt actatacttc tatagacaca caaacacaaa tacacacact aaagatctat
      301 gtctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
      361 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
      421 attgacctta aaattgattt gtactactgg taaattgcca gttccatggc caaccttagt
      481 cactacttta ggttatggtt tgcaatgttt tgctagatac ccagatcata tgaaacaaca
      541 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
      601 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
      661 tagaatcgaa ttaaaaggta ttgattttaa agaaggtggt aacattttag gtcacaaatt
      721 ggaatacaac tataactctc acaatgttta catcactgct gacaaacaaa agaatggtat
      781 caaagctaac ttcaaaatta gacacaacat tgaagatggt ggtgttcaat tagctgacca
      841 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
      901 atcctatcaa tctgccttat ccaaagatcc aaacgaaaag agagatcaca tggtcttgtt
      961 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aaggatccta
     1021 actcgagatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     1081 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     1141 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     1201 ttttgggacg ctcgaaggct gccaatgaga cgacggggtc atcacggctc atcatgcgcc
     1261 aaacaaatgt gtgcaataca cgctcggatg actgcatgat gaccgcactg actggggaca
     1321 gcagatccac ctaagcctgt gagagaagca gacacccgac agatcaaggc agttaactag
     1381 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1441 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1501 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1561 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1621 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1681 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1741 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     1801 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     1861 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     1921 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     1981 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     2041 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2101 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2161 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2221 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2281 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     2341 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     2401 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaga
     2461 gcacttgaat ccactgcccc gggaatctcg gtcgtaatga tttctataat gacgaaaaaa
     2521 aaaaaattgg aaagaaaaag cttcatggcc tttataaaaa ggaactatcc aatacctcgc
     2581 cagaaccaag taacagtatt ttacggggca caaatcaaga acaataagac aggactgtaa
     2641 agatggacgc attgaactcc aaagaacaac aagagttcca aaaagtagtg gaacaaaagc
     2701 aaatgaagga tttcatgcgt ttgtactcta atctggtaga aagatgtttc acagactgtg
     2761 tcaatgactt cacaacatca aagctaacca ataaggaaca aacatgcatc atgaagtgct
     2821 cagaaaagtt cttgaagcat agcgaacgtg tagggcagcg tttccaagaa caaaacgctg
     2881 ccttgggaca aggcttgggc cgataaggtg tactggcgta tatatatcta attatgtatc
     2941 tctggtgtag cccattttta gcatgtaaat ataaagaccg agcggccgcg attatcaaaa
     3001 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3061 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3121 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3181 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
     3241 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3301 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3361 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3421 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3481 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3541 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3601 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3661 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3721 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3781 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3841 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3901 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3961 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4021 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgtcatg
     4081 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4141 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4201 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4261 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4321 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4381 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4441 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4501 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4561 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4621 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4681 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4741 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4801 ttctttcctg cgttatcccc tgattctgtg gataaccgtg cggccgccaa tgctaaattc
     4861 gagtgaaaca caggaagatc agaaaatcct catttcatcc atattaacaa taatttcaaa
     4921 tgtttatttg cattatttga aactaggcaa gacaagcaac gaaacgtttt tgaaaatttt
     4981 gagtattttc aataaatttg tagaggactc agatattgaa aaaaagctac agcaattaat
     5041 acttgataag aagagtattg agaagggcaa cggttcatca tctcatggat ctgcacatga
     5101 acaaacacca gagtcaaacg acgttgaaat tgaggctact gcgccaattg atgacaatac
     5161 agacgatgat aacaaaccga agttatctga tgtagaaaag gattaaagat gctaagagat
     5221 agtgatgata tttcataaat aatgtaattc tatatatgtt aattaccttt tttgcgaggc
     5281 atatttatgg tgaaggataa gttttgacca tcaaagaagg ttaatgtggc tgtggtttca
     5341 gggtccataa agcccacatg gataacatta cccctgaatt cgcatctaga tggtagagcc
     5401 acaaacagcc ggtacaagca acgatctcca ggaccatctg aatcatgcgc ggatgacacg
     5461 aactcacgac ggcgatcaca gacattaacc cacagtacag acactgcgac aacgtggcaa
     5521 ttcgtcgcaa taccgtctca ctgaactggc cgataattgc agacg
//
