LOCUS       sequence-434477-        5551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene (2 operator) for zinc finger activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  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 5551)
  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: pKL1-423
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             285..998
                     /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      1014..1203
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             1558..2361
                     /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(3070..3861,3862..3930))
                     /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     3693..3712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3931..4035)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4138..4726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4627..4646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggaattc agacgctgct ctcgagacgc tgctcggatc ccagatccgc caggcgtgta
       61 tatatagcgt ggatggccag gcaactttag tgctgacaca tacaggcata tatatatgtg
      121 tgcgacgaca catgatcata tggcatgcat gtgctctgta tgtatataaa actcttgttt
      181 tcttcttttc tctaaatatt ctttccttat acattaggac ctttgcagca taaattacta
      241 tacttctata gacacacaaa cacaaataca cacactaaag atctatgtct aaaggtgaag
      301 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
      361 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
      421 tgatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt
      481 atggtttgca atgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt
      541 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
      601 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
      661 aaggtattga ttttaaagaa ggtggtaaca ttttaggtca caaattggaa tacaactata
      721 actctcacaa tgtttacatc actgctgaca aacaaaagaa tggtatcaaa gctaacttca
      781 aaattagaca caacattgaa gatggtggtg ttcaattagc tgaccattat caacaaaata
      841 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatctg
      901 ccttatccaa agatccaaac gaaaagagag atcacatggt cttgttagaa tttgttactg
      961 ctgctggtat tacccatggt atggatgaat tgtacaaagg atcctaactc gagatccgct
     1021 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     1081 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     1141 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     1201 aaggctgcca atgagacgac ggggtcatca cggctcatca tgcgccaaac aaatgtgtgc
     1261 aatacacgct cggatgactg catgatgacc gcactgactg gggacagcag atccacctaa
     1321 gcctgtgaga gaagcagaca cccgacagat caaggcagtt aactagtgca ctgcagtaca
     1381 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     1441 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     1501 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
     1561 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     1621 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     1681 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     1741 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     1801 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     1861 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     1921 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     1981 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     2041 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     2101 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     2161 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     2221 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     2281 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     2341 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     2401 caaattagag cttcaattta attatatcag ttattacccg agtagagcac ttgaatccac
     2461 tgccccggga atctcggtcg taatgatttc tataatgacg aaaaaaaaaa aattggaaag
     2521 aaaaagcttc atggccttta taaaaaggaa ctatccaata cctcgccaga accaagtaac
     2581 agtattttac ggggcacaaa tcaagaacaa taagacagga ctgtaaagat ggacgcattg
     2641 aactccaaag aacaacaaga gttccaaaaa gtagtggaac aaaagcaaat gaaggatttc
     2701 atgcgtttgt actctaatct ggtagaaaga tgtttcacag actgtgtcaa tgacttcaca
     2761 acatcaaagc taaccaataa ggaacaaaca tgcatcatga agtgctcaga aaagttcttg
     2821 aagcatagcg aacgtgtagg gcagcgtttc caagaacaaa acgctgcctt gggacaaggc
     2881 ttgggccgat aaggtgtact ggcgtatata tatctaatta tgtatctctg gtgtagccca
     2941 tttttagcat gtaaatataa agaccgagcg gccgcgatta tcaaaaagga tcttcaccta
     3001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3181 atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc cagatttatc
     3241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gtcatgacca aaatccctta
     4081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4801 atcccctgat tctgtggata accgtgcggc cgccaatgct aaattcgagt gaaacacagg
     4861 aagatcagaa aatcctcatt tcatccatat taacaataat ttcaaatgtt tatttgcatt
     4921 atttgaaact aggcaagaca agcaacgaaa cgtttttgaa aattttgagt attttcaata
     4981 aatttgtaga ggactcagat attgaaaaaa agctacagca attaatactt gataagaaga
     5041 gtattgagaa gggcaacggt tcatcatctc atggatctgc acatgaacaa acaccagagt
     5101 caaacgacgt tgaaattgag gctactgcgc caattgatga caatacagac gatgataaca
     5161 aaccgaagtt atctgatgta gaaaaggatt aaagatgcta agagatagtg atgatatttc
     5221 ataaataatg taattctata tatgttaatt accttttttg cgaggcatat ttatggtgaa
     5281 ggataagttt tgaccatcaa agaaggttaa tgtggctgtg gtttcagggt ccataaagcc
     5341 cacatggata acattacccc tgaattcgca tctagatggt agagccacaa acagccggta
     5401 caagcaacga tctccaggac catctgaatc atgcgcggat gacacgaact cacgacggcg
     5461 atcacagaca ttaacccaca gtacagacac tgcgacaacg tggcaattcg tcgcaatacc
     5521 gtctcactga actggccgat aattgcagac g
//
