LOCUS       sequence-472229-        4145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene VIII by Ptac promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4145)
  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 4145)
  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: pHK326
FEATURES             Location/Qualifiers
     source          1..4145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(220..1302)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1264..1283
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1423..1428,1429..1444,1445..1451)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1457..1476
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             1481..1503
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1510..1731
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     primer_bind     complement(2007..2025)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2021..2068
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2077..2163
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2255..2282
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2411..3202)
                     /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(3203..3294)
                     /gene="bla"
                     /label=AmpR promoter
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 cactgcccgc tttccagtcg
      241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      301 cgtattgggc gccagggtgg tttttctttt caccagtgag actggcaaca gctgattgcc
      361 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
      421 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctat cttcggtatc
      481 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
      541 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
      601 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
      661 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
      721 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
      781 cagatgctcc acgcccagtc gcgtaccgtc ctcatgggag aaaataatac tgttgatggg
      841 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
      901 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
      961 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     1021 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     1081 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     1141 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     1201 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     1261 accggcatac tctgcgacat cgtataacgt tactggtttc atattcacca ccctgaattg
     1321 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggcgcg
     1381 ccgccattgc ggccgcgccg tcactgcgtc ttttacgagc tgttgacaat taatcatcgg
     1441 ctcgtataat gtgtggaatt gtgagcgctc acaatttaat tttgtttaac tttaagaagg
     1501 agatatacca tgaaaaagtc tttagccctc aaagcctctg tagccgttgc taccctcgtt
     1561 ccgatgctgt ctttcgctgc tgagggtgac gatcccgcaa aagcggcctt taactccctg
     1621 caagcctcag cgaccgaata tatcggttat gcgtgggcga tggttgttgt cattgtcggc
     1681 gcaactatcg gtatcaagct gtttaagaaa ttcacctcga aagcaagctg actcggtacc
     1741 aaattccaga aaagaggcct cccgaaaggg gggccttttt tcgttttggt cctaatagat
     1801 aaaggatagg tctggtagtg ttgttcgttc tcgcaggtaa atcaataata ctcagcagtt
     1861 ccgtagactt ttcagtggga cagggtagcg ataacagata gattgtaata agacacagta
     1921 ggtgctcgta gttgcgtgaa gagaaccgct caggaaatcc agtcagaagt attggtaatc
     1981 gttgaaaact cagtcgactg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2041 ctctaaacgg gtcttgaggg gttttttgcc aggcatcaaa taaaacgaaa ggctcagtcg
     2101 gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     2161 aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga
     2221 cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatcctgac ggatggcctt
     2281 tttgcgtttc agatctaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     2341 gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa
     2401 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
     2461 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
     2521 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
     2581 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
     2641 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
     2701 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     2761 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
     2821 cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt
     2881 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     2941 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     3001 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     3061 tatcactgtg tggcttcagg ccgtcatcca ctgcggagcc gtacaaatgt acggccagca
     3121 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     3181 cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg
     3241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta
     3301 gctcactcgg tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa
     3361 agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc
     3421 cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg
     3481 cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga
     3541 aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc
     3601 ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt
     3661 ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt ttgctccaag
     3721 ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt
     3781 tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac
     3841 tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa
     3901 agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac
     3961 ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt
     4021 tcgaaggtta agtcagttgg ggaactgctt aaccttcgat caaaccacct ccccaggtgg
     4081 ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4141 tgatc
//
