LOCUS       sequence-434483-        5537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene (1 operator) for zinc finger activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  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 5537)
  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: pKL1-422
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             271..984
                     /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      1000..1189
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             1544..2347
                     /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(3056..3847,3848..3916))
                     /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     3679..3698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3917..4021)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4124..4712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4613..4632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacggaattc agacgctgct cggatcccag atccgccagg cgtgtatata tagcgtggat
       61 ggccaggcaa ctttagtgct gacacataca ggcatatata tatgtgtgcg acgacacatg
      121 atcatatggc atgcatgtgc tctgtatgta tataaaactc ttgttttctt cttttctcta
      181 aatattcttt ccttatacat taggaccttt gcagcataaa ttactatact tctatagaca
      241 cacaaacaca aatacacaca ctaaagatct atgtctaaag gtgaagaatt attcactggt
      301 gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc
      361 ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaattgat ttgtactact
      421 ggtaaattgc cagttccatg gccaacctta gtcactactt taggttatgg tttgcaatgt
      481 tttgctagat acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa
      541 ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct
      601 gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt
      661 aaagaaggtg gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt
      721 tacatcactg ctgacaaaca aaagaatggt atcaaagcta acttcaaaat tagacacaac
      781 attgaagatg gtggtgttca attagctgac cattatcaac aaaatactcc aattggtgat
      841 ggtccagtct tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat
      901 ccaaacgaaa agagagatca catggtcttg ttagaatttg ttactgctgc tggtattacc
      961 catggtatgg atgaattgta caaaggatcc taactcgaga tccgctctaa ccgaaaagga
     1021 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     1081 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     1141 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctgccaatga
     1201 gacgacgggg tcatcacggc tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga
     1261 tgactgcatg atgaccgcac tgactgggga cagcagatcc acctaagcct gtgagagaag
     1321 cagacacccg acagatcaag gcagttaact agtgcactgc agtacacggt ttccttgaaa
     1381 tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag
     1441 attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac
     1501 ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata
     1561 taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca
     1621 cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga
     1681 gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac
     1741 tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt
     1801 tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc
     1861 tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg
     1921 cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg
     1981 ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa
     2041 gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag
     2101 agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt
     2161 agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac
     2221 aggatctgac attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt
     2281 agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca
     2341 aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc
     2401 aatttaatta tatcagttat tacccgagta gagcacttga atccactgcc ccgggaatct
     2461 cggtcgtaat gatttctata atgacgaaaa aaaaaaaatt ggaaagaaaa agcttcatgg
     2521 cctttataaa aaggaactat ccaatacctc gccagaacca agtaacagta ttttacgggg
     2581 cacaaatcaa gaacaataag acaggactgt aaagatggac gcattgaact ccaaagaaca
     2641 acaagagttc caaaaagtag tggaacaaaa gcaaatgaag gatttcatgc gtttgtactc
     2701 taatctggta gaaagatgtt tcacagactg tgtcaatgac ttcacaacat caaagctaac
     2761 caataaggaa caaacatgca tcatgaagtg ctcagaaaag ttcttgaagc atagcgaacg
     2821 tgtagggcag cgtttccaag aacaaaacgc tgccttggga caaggcttgg gccgataagg
     2881 tgtactggcg tatatatatc taattatgta tctctggtgt agcccatttt tagcatgtaa
     2941 atataaagac cgagcggccg cgattatcaa aaaggatctt cacctagatc cttttaaatt
     3001 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3061 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3121 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3181 ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3241 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3481 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3721 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3901 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3961 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4021 gcacatttcc ccgaaaagtg ccacctgtca tgaccaaaat cccttaacgt gagttttcgt
     4081 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4141 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4201 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4261 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4321 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4381 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4441 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4501 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4561 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4621 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4681 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4741 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4801 tggataaccg tgcggccgcc aatgctaaat tcgagtgaaa cacaggaaga tcagaaaatc
     4861 ctcatttcat ccatattaac aataatttca aatgtttatt tgcattattt gaaactaggc
     4921 aagacaagca acgaaacgtt tttgaaaatt ttgagtattt tcaataaatt tgtagaggac
     4981 tcagatattg aaaaaaagct acagcaatta atacttgata agaagagtat tgagaagggc
     5041 aacggttcat catctcatgg atctgcacat gaacaaacac cagagtcaaa cgacgttgaa
     5101 attgaggcta ctgcgccaat tgatgacaat acagacgatg ataacaaacc gaagttatct
     5161 gatgtagaaa aggattaaag atgctaagag atagtgatga tatttcataa ataatgtaat
     5221 tctatatatg ttaattacct tttttgcgag gcatatttat ggtgaaggat aagttttgac
     5281 catcaaagaa ggttaatgtg gctgtggttt cagggtccat aaagcccaca tggataacat
     5341 tacccctgaa ttcgcatcta gatggtagag ccacaaacag ccggtacaag caacgatctc
     5401 caggaccatc tgaatcatgc gcggatgaca cgaactcacg acggcgatca cagacattaa
     5461 cccacagtac agacactgcg acaacgtggc aattcgtcgc aataccgtct cactgaactg
     5521 gccgataatt gcagacg
//
