LOCUS       sequence-472226-        4686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene cinI for OC14-HSL production by Ptac promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4686)
  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 4686)
  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: pHK-cinI
FEATURES             Location/Qualifiers
     source          1..4686
                     /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(1438..1443,1444..1459,1460..1466)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1472..1491
                     /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."
     protein_bind    1552..1566
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease 
                     (Haurwitz et al., 2010)"
     RBS             1567..1578
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2303..2322
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     primer_bind     complement(2613..2631)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2627..2674
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2683..2769
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2861..2888
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3017..3808)
                     /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(3809..3900)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3948..4686)
                     /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 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 tcagcctgcg gtccgggttg
     1441 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cgctcacaat ttcgctggga
     1501 cgcccgagag ccgaatcgca cttatttaca gtagttcagc ctgcggtccg gctgccgtat
     1561 aggcagaaag aggagaaaat gcgggggagt gttactagta tgttcgttat cattcaggca
     1621 catgagtatc agaaatacgc tgccgtactc gaccagatgt ttcgtctgcg caagaaggtc
     1681 ttcgccgata cgctctgctg ggacgttcct gtcatcggcc cttacgaacg tgacagctac
     1741 gattcgcttg ctcccgccta tctcgtctgg tgcaacgaca gccgcacccg tctttatggc
     1801 ggcatgcgcc tgatgccgac gaccggcccg acccttctct acgacgtctt ccgcgagacg
     1861 ttccctgatg ccgccgatct tatcgccccc ggcatctggg aaggcacgcg catgtgcatc
     1921 gacgaggagg cgatcgccaa ggatttcccc gagatcgacg ccggccgcgc cttctccatg
     1981 atgctgctcg cgctttgcga atgcgcgctc gatcacggca tccacacgat gatctccaac
     2041 tacgagccct acctcaagcg cgtctacaag cgcgccggcg ccgaggtgga agaactcggc
     2101 cgcgcagacg gctacggcaa atatcccgtc tgctgcggcg ccttcgaagt ctcggaccgc
     2161 gtgctgcgca agatgcgcgc cgccctcggc ctcaccctac ccctttatgt caggcacgtg
     2221 ccggcccgct cggtcgtgac ccaattcctg gagatggcag cagcagcgaa cgacgaaaat
     2281 tacgcccttg cagcgtaata aagttcactg ccgtataggc agtcgctggg acgcccgctc
     2341 ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt tttggtccta
     2401 atagataaag gataggtctg gtagtgttgt tcgttctcgc aggtaaatca ataatactca
     2461 gcagttccgt agacttttca gtgggacagg gtagcgataa cagatagatt gtaataagac
     2521 acagtaggtg ctcgtagttg cgtgaagaga accgctcagg aaatccagtc agaagtattg
     2581 gtaatcgttg aaaactcagt cgactgctgc caccgctgag caataactag cataacccct
     2641 tggggcctct aaacgggtct tgaggggttt tttgccaggc atcaaataaa acgaaaggct
     2701 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     2761 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
     2821 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
     2881 ggcctttttg cgtttcagat ctaccggtaa accagcaata gacataagcg gctatttaac
     2941 gaccctgccc tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg
     3001 aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
     3061 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
     3121 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     3181 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     3241 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     3301 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     3361 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     3421 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     3481 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     3541 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     3601 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
     3661 aatcaatatc actgtgtggc ttcaggccgt catccactgc ggagccgtac aaatgtacgg
     3721 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
     3781 atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt
     3841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3901 tagctagctc actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac
     3961 atacaaagtt acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag
     4021 cagggccgcg ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa
     4081 cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac
     4141 gggcgaaacc cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc
     4201 gctctcctgt tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg
     4261 aagtgtggcg ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgtttgc
     4321 tccaagctgg gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt
     4381 aactgttcac ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt
     4441 ggtaactggg agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg
     4501 cgccaaagtc cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt
     4561 taccacggtt aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg
     4621 gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct
     4681 ttgatc
//
