LOCUS       sequence-472227-        3831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene VIII by Plux promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3831)
  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 3831)
  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: pHK336
FEATURES             Location/Qualifiers
     source          1..3831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1232..1254
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1261..1482
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     primer_bind     complement(1758..1776)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1772..1819
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1828..1914
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2006..2033
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2162..2953)
                     /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(2954..3045)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3093..3831)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
       61 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
      121 aggagctgac tgggttgctc ctaggatgca gtgactcgca catttccccg aaaagtgcca
      181 cctgaattcg aaatccctga aactgagact gtagaaaatt tattaatttt tgaagtacgg
      241 acagtcgata gcgccggtca ggattgcttt ggagatggat tggcagcgat tcgtggtgtt
      301 cagtttcatt tgcgcgttcg tcagatgaaa cgtaacggta cgttcggaac aacccaggat
      361 tttactaata tcccaagagc ttttgccttc acaggcccac gccaggcatt ctttttcgcg
      421 tttggtcagg tcattgttac ttttgttgtt agcgatgttg attttacggt agttatcaac
      481 cagagacggc acgatcagcg gaatattcat gcatgcatgc aggaacaggc tgtcaatata
      541 gttatctttt tcagagtgag cgaagctcag cataccaaag ccattgtttg cggtatggat
      601 cgggaacgaa aaacccgtaa tcaggccgct ggttttggct tctttgatca cgttcggaga
      661 ttttttgttg accgcattgt tttcaaagat attccagtta atcggggagt gattggagtt
      721 actgtagtcc acgatcggat catatttaat cagattggcg tcatcgtaat actgacgcca
      781 ttttttcggg tagttgtcca ggatcgaaat atctgatttg accatactgt gcggatagat
      841 aatcgccagc aggtaatatt cgcaatgaac cattttggtc atatcgctca ggcactggtt
      901 gatgtcgttg tttgagcgac aagctttgat tttgttgatg atgcggtacg tatcgtcggc
      961 gttgatgttt ttcataccca tctctttatc cttacctatt gtttgtcgca agttttgcgt
     1021 gttatatatc attaaaacgg taatggattg acatttgatt ctaataaatt ggatttttgt
     1081 cacactattg tatcgctggg aatacaatta cttaacataa gcacctgtag gatcgtacag
     1141 gtcaataaac agttgatagg gcttctccgt taacctgtag gatcgtacag gtttacgcaa
     1201 gaaaatggtt tgttatagtc gaataaataa ttttgtttaa ctttaagaag gagatatacc
     1261 atgaaaaagt ctttagccct caaagcctct gtagccgttg ctaccctcgt tccgatgctg
     1321 tctttcgctg ctgagggtga cgatcccgca aaagcggcct ttaactccct gcaagcctca
     1381 gcgaccgaat atatcggtta tgcgtgggcg atggttgttg tcattgtcgg cgcaactatc
     1441 ggtatcaagc tgtttaagaa attcacctcg aaagcaagct gactcggtac caaattccag
     1501 aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tcctaataga taaaggatag
     1561 gtctggtagt gttgttcgtt ctcgcaggta aatcaataat actcagcagt tccgtagact
     1621 tttcagtggg acagggtagc gataacagat agattgtaat aagacacagt aggtgctcgt
     1681 agttgcgtga agagaaccgc tcaggaaatc cagtcagaag tattggtaat cgttgaaaac
     1741 tcagtcgact gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1801 ggtcttgagg ggttttttgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
     1861 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1921 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     1981 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
     2041 cagatctacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     2101 cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac
     2161 attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact
     2221 gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa
     2281 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     2341 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     2401 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     2461 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     2521 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     2581 ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     2641 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     2701 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     2761 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     2821 gtggcttcag gccgtcatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt
     2881 cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca
     2941 ccgcttccct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3001 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg
     3061 gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca
     3121 cagattccgt ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt
     3181 ggcgtttttc cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg
     3241 tgcatctgtg ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca
     3301 ggacttaaag atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga
     3361 ccctgccgtt taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc
     3421 tcatagctca cacactggta tctcggctcg gtgtaggtcg tttgctccaa gctgggctgt
     3481 aagcaagaac tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag
     3541 tccaacccgg aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc
     3601 gcagaggatt tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct
     3661 acactggaag gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca
     3721 gttccccaac tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca
     3781 gggcaaaaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat c
//
