LOCUS       sequence-385742-        3097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cCadC-GCN4* under the control of Ptet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3097)
  AUTHORS   Lee B, Wang T
  TITLE     A Modular Scaffold for Controlling Transcriptional Activation via 
            Homomeric Protein-Protein Interactions.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3198-3206. doi: 
            10.1021/acssynbio.2c00501. Epub 2022 Oct 10.
  PUBMED    36215660
REFERENCE   2  (bases 1 to 3097)
  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: 
            "10.1021/acssynbio.2c00501"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3198-3206"
COMMENT     Sequence Label: pBL069B
FEATURES             Location/Qualifiers
     source          1..3097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    782..922
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(848..867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1008..1030
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1024..1215)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1412..1430
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1431..1485
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1436..1454
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1466..1484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2065..2130
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             complement(2213..3004)
                     /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"
     promoter        complement(3005..3096)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taggccgttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc acctcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      901 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      961 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1021 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1081 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     1141 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     1201 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     1261 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac aagaggacat
     1321 ccggtcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt gctgaggaga
     1381 cttagggacc ctccatcgaa tggccagatg atccctatca gtgatagaga ttgacactct
     1441 atcattgata gagttatttt accactccct atcagtgata gagaaaagtg aaatgaatag
     1501 ttcgacaaaa atctagacct gcaggtgcag taaaggaaaa aaaaaatgca acaacctgta
     1561 gttcgcgttg gcgaatggct tgttactccg tccataaacc aaattagccg caatgggcgt
     1621 caacttaccc ttcgtccgag attaatcgat cttctggttt tctttgctca acacagtggc
     1681 gaagtactta gcagggatga acttatcgat aatgtctgga agagaagtat tgtcaccaat
     1741 cacgttgtga cgcagagtat ctcagaacta cgtaagtcat taaaagataa tgatgaagat
     1801 agtcctgtct atatcgctac tgtaccaaag cgcggctata aattaatggt gccggttatc
     1861 tggtacagcg aagaagaggg agaggaaata atgctatctt cggcagcggc tatctctgag
     1921 gcggttcctg ccacagattc tccctcccac agtcttaaca ttcaaaacac cgcaacgcca
     1981 cctgaacaat ccccagttaa aagcaaacga ggtggatctg gtggtttgca aagaatgaaa
     2041 caacttgaac cgaaggttga agaattgctt ccgaaaaatt atcacttgga aaatgaggtt
     2101 gccagattaa agaaattagt tggcgaacgc taaacttaat taacggcact cctcagcaaa
     2161 tataatgacc ctcttgataa cccaagaggg cattttttaa tgcccatggc gtttatttgc
     2221 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     2281 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     2341 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     2401 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     2461 ctccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     2521 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     2581 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     2641 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     2701 gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa
     2761 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     2821 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     2881 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     2941 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     3001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3061 gatacatatt tgaatgtatt tagaaaaata ggccaaa
//
