LOCUS       sequence-472228-        4016 bp    DNA     circular SYN 09-MAY-2025
DEFINITION  Inducible gene VIII by Pbad promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4016)
  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 4016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 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: pHK346
FEATURES             Location/Qualifiers
     source          1..4016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(220..1098)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1125..1409
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1332..1351
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1417..1439
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1446..1667
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     primer_bind     complement(1943..1961)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1957..2004
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2013..2099
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2191..2218
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2347..3138)
                     /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(3139..3230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3278..4016)
                     /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 tatgacaact tgacggctac
      241 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
      301 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
      361 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
      421 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
      481 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
      541 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
      601 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
      661 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
      721 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
      781 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
      841 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
      901 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
      961 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     1021 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     1081 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     1141 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     1201 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     1261 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     1321 cacactttgc tatgccatag catttttatc cataagatta gcggttccta cctgacgctt
     1381 tttatcgcaa ctctctactg tttctccata cctaattttg tttaacttta agaaggagat
     1441 ataccatgaa aaagtcttta gccctcaaag cctctgtagc cgttgctacc ctcgttccga
     1501 tgctgtcttt cgctgctgag ggtgacgatc ccgcaaaagc ggcctttaac tccctgcaag
     1561 cctcagcgac cgaatatatc ggttatgcgt gggcgatggt tgttgtcatt gtcggcgcaa
     1621 ctatcggtat caagctgttt aagaaattca cctcgaaagc aagctgactc ggtaccaaat
     1681 tccagaaaag aggcctcccg aaaggggggc cttttttcgt tttggtccta atagataaag
     1741 gataggtctg gtagtgttgt tcgttctcgc aggtaaatca ataatactca gcagttccgt
     1801 agacttttca gtgggacagg gtagcgataa cagatagatt gtaataagac acagtaggtg
     1861 ctcgtagttg cgtgaagaga accgctcagg aaatccagtc agaagtattg gtaatcgttg
     1921 aaaactcagt cgactgctgc caccgctgag caataactag cataacccct tggggcctct
     1981 aaacgggtct tgaggggttt tttgccaggc atcaaataaa acgaaaggct cagtcggaag
     2041 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2101 cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc
     2161 cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat ggcctttttg
     2221 cgtttcagat ctaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     2281 tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg aacgaattgt
     2341 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     2401 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     2461 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     2521 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     2581 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     2641 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     2701 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     2761 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     2821 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     2881 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     2941 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     3001 actgtgtggc ttcaggccgt catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     3061 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     3121 gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagctagctc
     3241 actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac atacaaagtt
     3301 acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag cagggccgcg
     3361 ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa cagacgcttt
     3421 tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc
     3481 cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc gctctcctgt
     3541 tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg aagtgtggcg
     3601 ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgtttgc tccaagctgg
     3661 gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt aactgttcac
     3721 ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt ggtaactggg
     3781 agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg cgccaaagtc
     3841 cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt taccacggtt
     3901 aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg gttttttcgt
     3961 ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct ttgatc
//
