LOCUS       sequence-422172-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mNeonGreen and mCherry from aTc-inducible Ptet 
            promoters. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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: pPtet-1_mNeonGreen_Ptet-2_mCherry
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    82..100
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    121..139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1174..1192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1213..1231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2450..2637
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2638..3300
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     misc_feature    3515..4018
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4035..4826)
                     /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      5256..5844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5745..5764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagac tgaactggcc gataattgca gacgaacgtt tttttctttt
       61 tttttttttg cggtataaaa atccctatca gtgatagaga ttttgttttt tttttttttt
      121 tccctatcag tgatagagaa tatcaagcta caaaaaggta tggtttcaaa gggtgaagag
      181 gataacatgg caagtttgcc ggccacacat gaacttcaca ttttcgggag cataaatggc
      241 gttgattttg acatggtcgg tcaggggact gggaacccca acgacggata cgaagaactc
      301 aacttgaaat ctaccaaagg tgatctccaa ttttcaccgt ggattcttgt tccgcacatt
      361 gggtatggat ttcatcagta tttgccatac cctgatggca tgtcaccctt ccaggccgct
      421 atggtcgatg gtagcggcta ccaggtacac cgcacaatgc agtttgaaga cggtgcatca
      481 ctgaccgtga actaccgata tacctatgag ggttcccata ttaaaggaga agctcaagta
      541 aagggaacgg gattcccggc cgatggtccc gtgatgacta actccttgac agcagcagac
      601 tggtgtcgct ctaagaagac ttatcccaac gataaaacca ttataagtac atttaaatgg
      661 agttatacaa cgggcaacgg taagaggtac agaagcacgg cacggaccac ttacaccttc
      721 gctaagccga tggccgcaaa ctacttgaag aaccaaccga tgtacgtatt ccgaaagacg
      781 gaactgaaac atagcaaaac tgaactgaat ttcaaggagt ggcagaaagc gtttactgat
      841 gtcatgggta tggacgagct gtataaggga tcctaactcg agagtgcttt taactaagaa
      901 ttattagtct tttctgctta ttttttcatc atagtttaga acactttata ttaacgaata
      961 gtttatgaat ctatttaggt ttaaaaattg atacagtttt ataagttact ttttcaaaga
     1021 ctcgtgctgt ctattgcata atgcactgga aggggaaaaa aaaggtgcac acgcgtggct
     1081 ttttcttgaa tttgcagttt gaaaaatgct gccaaaccag atgtcaacac agctacaacg
     1141 tttttttatt ttttgttttt tgcggtataa aaatccctat cagtgataga gattttattt
     1201 tttttttctt tttccctatc agtgatagag aatatcaagc tacaaaaagg tatggtatct
     1261 aaaggagaag aggataacat ggcgataatt aaggagttta tgagatttaa ggtgcatatg
     1321 gaaggcagcg ttaatggaca tgagttcgaa atcgaaggtg aaggcgaggg gcgtccctat
     1381 gagggaacac aaaccgcaaa actgaaagtt accaaaggcg gaccccttcc ttttgcctgg
     1441 gatatactgt caccacaatt catgtatggc tccaaggctt atgtcaaaca tcccgcagac
     1501 attcccgatt acctaaaact tagcttcccg gaaggcttta agtgggaaag ggttatgaat
     1561 ttcgaagacg ggggcgtggt cactgtgacc caggacagta gtcttcagga tggagagttc
     1621 atatacaaag ttaaactgcg tggaaccaat ttcccttcag acggaccagt gatgcagaag
     1681 aaaaccatgg gatgggaggc gtcctctgaa agaatgtatc ccgaagacgg tgccctaaag
     1741 ggtgagatca agcagagact aaagttaaag gatggcggac attatgacgc cgaggtaaag
     1801 acgacttata aggcgaagaa acccgtgcag ctgccagggg cttacaatgt gaatataaaa
     1861 ttagatatta ccagccataa cgaagattat acgatagttg aacagtacga gagagcggaa
     1921 ggtcgtcaca gtaccggggg catggacgaa ttgtataaag gatcctaaat aaagcaatct
     1981 tgatgaggat aatgattttt ttttgaatat acataaatac taccgttttt ctgctagatt
     2041 ttgtgatgac gtaaataagt acatattact ttttaagcca agacaagatt aagcattaac
     2101 tttacccttt tctttctaag tttcaatatt agttatcact gtttaaaagt tatggcgaga
     2161 acgtcggcgg ttaaaatata ttaccctgaa cgcctcttcc agcgctcaac gggtgtgctt
     2221 cccgttctga tgagtccgtg aggacgaaag cgtcccggga atatcgcgta gcactttaga
     2281 ctcttcctta cgtaatggat gactcggcgc gtgatcttac ggccgggcga cacctctcat
     2341 aggtgtgcgt gggctgagca gttacagaga tgttacgaac cactagtgca ctgcagtaca
     2401 aacacagtcc tttcccgcaa ttttcttttt ctattactct tggcctcctc tagtacactc
     2461 tatatttttt tatgcctcgg taatgatttt catttttttt tttccaccta gcggatgact
     2521 cttttttttt cttagcgatt ggcattatca cataatgaat tatacattat ataaagtaat
     2581 gtgatttctt cgaagaatat actaaaaaat gagcaggcaa gataaacgaa ggcaaagatg
     2641 acagagcaga aagccctagt aaagcgtatt acaaatgaaa ccaagattca gattgcgatc
     2701 tctttaaagg gtggtcccct agcgatagag cactcgatct tcccagaaaa agaggcagaa
     2761 gcagtagcag aacaggccac acaatcgcaa gtgattaacg tccacacagg tatagggttt
     2821 ctggaccata tgatacatgc tctggccaag cattccggct ggtcgctaat cgttgagtgc
     2881 attggtgact tacacataga cgaccatcac accactgagg actgcgggat tgctctcggt
     2941 caagctttta aagaggccct aggggccgtg cgtggagtaa aaaggtttgg atcaggattt
     3001 gcgcctttgg atgaggcact ttccagagcg gtggttgatc tttcgaacag gccgtacgca
     3061 gttgtcgaac ttggtttgca aagggagaaa gtaggtgatc tctcttgcga gatgatcccg
     3121 cattttcttg aaagctttgc agaggctagc agaattaccc tccacgttga ttgtctgcga
     3181 ggcaagaatg atcatcaccg tagtgagagt gcgttcaagg ctcttgcggt tgccataaga
     3241 gaagccacct cgcccaatgg taccaacgat gttccctcca ccaaaggtgt tcttatgtag
     3301 tgacaccgat tatttaaagc tgctgcatac gatatatata catgtgtata tatgtatacc
     3361 tatgaatgtc agtaagtatg tatacgaaca gtatgatact gaagatgaca aggtaatgca
     3421 tcattctata cgtgtcattc tgaacgaggc gcgctttcct tttttctttt tgctttttct
     3481 ttttttttct cttgaactcg acggatcata gagtatcacg tgctataaaa ataattataa
     3541 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     3601 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     3661 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     3721 gtgtagaaaa ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     3781 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     3841 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     3901 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     3961 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg
     4021 tcccgagcgg ccgcttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     4081 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     4141 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     4201 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     4261 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     4321 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     4381 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     4441 atgtcgatcg tggctggctc gaagataccc gcaagaatgt cattgcgctg ccattctcca
     4501 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     4561 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     4621 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     4681 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     4741 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     4801 acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg actgccctgc
     4861 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     4921 tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca agccatgaaa
     4981 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     5041 cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca ctgggttcgt
     5101 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag
     5161 gcatttctgt cctggctggt catgaccaaa atcccttaac gtgagttttc gttccactga
     5221 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5341 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5401 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5941 cgtgcggccg c
//
