LOCUS       sequence-385743-        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: pBL069
FEATURES             Location/Qualifiers
     source          1..3097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    776..916
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(842..861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1002..1024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1018..1209)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1406..1424
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1425..1479
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1430..1448
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1460..1478
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2053..2109
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             complement(2207..2998)
                     /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(2999..3090)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccacctct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
      781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      841 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
      901 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      961 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1021 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     1081 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     1141 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     1201 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     1261 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacaagagg acatccggtc
     1321 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttgctgag gagacttagg
     1381 gaccctccat cgaatggcca gatgatccct atcagtgata gagattgaca ctctatcatt
     1441 gatagagtta ttttaccact ccctatcagt gatagagaaa agtgaaatga atagttcgac
     1501 aaaaatctag acctgcaggt gcagtaaagg aaaaaaaaaa tgcaacaacc tgtagttcgc
     1561 gttggcgaat ggcttgttac tccgtccata aaccaaatta gccgcaatgg gcgtcaactt
     1621 acccttcgtc cgagattaat cgatcttctg gttttctttg ctcaacacag tggcgaagta
     1681 cttagcaggg atgaacttat cgataatgtc tggaagagaa gtattgtcac caatcacgtt
     1741 gtgacgcaga gtatctcaga actacgtaag tcattaaaag ataatgatga agatagtcct
     1801 gtctatatcg ctactgtacc aaagcgcggc tataaattaa tggtgccggt tatctggtac
     1861 agcgaagaag agggagagga aataatgcta tcttcggcag cggctatctc tgaggcggtt
     1921 cctgccacag attctccctc ccacagtctt aacattcaaa acaccgcaac gccacctgaa
     1981 caatccccag ttaaaagcaa acgaggtgga tctggtggtt tgcaaagaat gaaacaactt
     2041 gaagacaagg ttgaagaatt gctttcgaaa aattatcact tggaaaatga ggttgccaga
     2101 ttaaagaaat tagttggcga acgctaaact taattaacgg cactcctcag caaatataat
     2161 gaccctcttg ataacccaag agggcatttt ttaatgccca tggcgtttat ttgccgacta
     2221 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     2281 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     2341 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     2401 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatctccag
     2461 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     2521 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     2581 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     2641 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     2701 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     2761 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     2821 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     2881 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     2941 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac
     3001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3061 tatttgaatg tatttagaaa aataggccaa ataggcc
//
