LOCUS       sequence-472231-        3759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene VIII by Ptet promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3759)
  AUTHORS   Kusumawardhani, H., Zoppi, F., Avendano, R. et al.
  TITLE     Engineering intercellular communication using M13 phagemid and 
            CRISPR-based gene regulation for multicellular computing in 
            Escherichia coli
  JOURNAL   Nat Commun 16, 3569 (2025)
REFERENCE   2  (bases 1 to 3759)
  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"; journalName: "Nat Commun 
            16, 3569 (2025)"
COMMENT     Sequence Label: pHK316
FEATURES             Location/Qualifiers
     source          1..3759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1024..1647)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1555..1574
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     1804..1821
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1805..1821
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1841..1859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1866..1884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1899..1921
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1928..2149
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     primer_bind     complement(2425..2443)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2439..2486
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2495..2581
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2673..2700
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2829..3620)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDDGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3621..3712)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
       61 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
      121 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
      181 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
      241 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
      301 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
      361 gtaggtcgtt tgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
      421 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
      481 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
      541 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
      601 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
      661 acctccccag gtggtttttt cgaaggttaa gtcagttggg gaactgctta accttcgatc
      721 aaaccacctc cccaggtggt tttttcgttt acagggcaaa agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct actgaaccgc tctagatttc agtgcaattt
      841 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
      901 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gctcctagga tgcagtgact
      961 cgcacatttc cccgaaaagt gccacctgaa ttcgaaatcc ctgaaactga gactgtagaa
     1021 aatttaggac ccactttcac atttaagttg tttttctaat ccgcatataa tcaattcaag
     1081 gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa
     1141 taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc
     1201 ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc
     1261 attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata
     1321 ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa
     1381 agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa
     1441 agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc
     1501 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag
     1561 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat
     1621 cactttactt ttatctaatc tggacatttt tttcctcctg gatccttaat ttttaaagta
     1681 tgggcaatca attgctcctg ttaaaattgc tttagaaata ctttggcagc ggtttgttgt
     1741 attgagtttc atttgcgcat tggttaaatg gaaagtgaca gtacgctcac tgcgaattcg
     1801 agttgtaaaa cgacggccag tactccaccg ttggcttttt tccctatcag tgatagagat
     1861 tgacatccct atcagtgata gagataatga gcactaattt tgtttaactt taagaaggag
     1921 atataccatg aaaaagtctt tagccctcaa agcctctgta gccgttgcta ccctcgttcc
     1981 gatgctgtct ttcgctgctg agggtgacga tcccgcaaaa gcggccttta actccctgca
     2041 agcctcagcg accgaatata tcggttatgc gtgggcgatg gttgttgtca ttgtcggcgc
     2101 aactatcggt atcaagctgt ttaagaaatt cacctcgaaa gcaagctgac tcggtaccaa
     2161 attccagaaa agaggcctcc cgaaaggggg gccttttttc gttttggtcc taatagataa
     2221 aggataggtc tggtagtgtt gttcgttctc gcaggtaaat caataatact cagcagttcc
     2281 gtagactttt cagtgggaca gggtagcgat aacagataga ttgtaataag acacagtagg
     2341 tgctcgtagt tgcgtgaaga gaaccgctca ggaaatccag tcagaagtat tggtaatcgt
     2401 tgaaaactca gtcgactgct gccaccgctg agcaataact agcataaccc cttggggcct
     2461 ctaaacgggt cttgaggggt tttttgccag gcatcaaata aaacgaaagg ctcagtcgga
     2521 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2581 tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc gggcaggacg
     2641 cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt
     2701 tgcgtttcag atctaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     2761 cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt
     2821 gttagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     2881 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     2941 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     3001 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     3061 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     3121 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     3181 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     3241 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     3301 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     3361 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     3421 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     3481 tcactgtgtg gcttcaggcc gtcatccact gcggagccgt acaaatgtac ggccagcaac
     3541 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     3601 gcgatcaccg cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc
     3721 tcactcggtc gctacgctcc gggcgtgaga ctgcggcgg
//
