LOCUS       sequence-455034-        8725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kill switch plasmid for DNA sensor construction using Bacillus 
            subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8725)
  AUTHORS   Cheng YY, Chen Z, Cao X, Ross TD, Falbel TG, Burton BM, Venturelli 
            OS
  TITLE     Programming bacteria for multiplexed DNA detection.
  JOURNAL   Nat Commun. 2023 Apr 10;14(1):2001. doi: 10.1038/s41467-023-37582-x.
  PUBMED    37037805
REFERENCE   2  (bases 1 to 8725)
  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."; date: "2023-04-10"; volume: "14(1):2001. doi"; pages: " 
            10.1038/s41467-023-37582-x"
COMMENT     Sequence Label: pOSV00157
FEATURES             Location/Qualifiers
     source          1..8725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(808..902)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(954..2036)
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1998..2017
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    3005..3021
                     /label=lac operator
                     /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)."
     terminator      complement(3125..3159)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     terminator      complement(3239..3266)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3285..3371)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(6896..7484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6976..6995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7655..8446,8447..8515))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8278..8297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8516..8620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8688..8706
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
       61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
      121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
      181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
      241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
      301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
      361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
      421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
      481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
      541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
      601 catggaaaca cacaaattaa aaactggtct gatcgatggg atgtcacgca gaattaaata
      661 gtacataatg gatttcctta cgcgaaatac gggcagacat ggcctgcccg gttattatta
      721 tttttgacac cagaccaact ggtaatgcga gaccggtctc ggtagcgacc ggcgctcagg
      781 atccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
      841 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
      901 tccaagcgag ctcgatatca aattacgcct aactcacatt aattgcgttg cgctcactgc
      961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1021 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     1081 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     1141 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     1201 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     1261 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     1321 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     1381 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     1441 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     1501 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     1561 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     1621 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     1681 acgcgttgcg cgagaagatt gtgcaccgcc gttttacagg cttcgacgcc gcttcgttct
     1741 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     1801 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     1861 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     1921 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     1981 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcatcaaa
     2041 atcgtctccc tccgtttgaa tatttgattg atcgtaacca gatgaagcac tctttccact
     2101 atccctacag tgttatggct tgaacaatca cgaaacaata attggtacgt acgatctttc
     2161 agccgactca aacatcaaat cttacaaatg tagtctttga aagtattaca tatgtaagat
     2221 ttaaatgcaa ccgttttttc ggaaggaaat gatggcctcg tttccaccga attagcttgc
     2281 atgcggctag cactttaccc ttttactttg ccgagctcaa ggggtgggtc tatttcttct
     2341 ttatataagt caacagggca attataaatg acccgaaagc aagcataagc attaatgctt
     2401 gaaatgttga cacgtgcatc accccctttc atgagggagt gagccagacc gcccttgaga
     2461 tgttgaaagc caatattcat ttatactaaa ccatattgta ccataacact ttcataatcc
     2521 gagggttttt aataaaagaa aaagggttaa ggcttctatg atcaggtcat tttgaactca
     2581 tagataaaaa aagtaacaaa taaaagatac aaataattat ataatcaata tatattcatg
     2641 ttataatggt gccaagcaaa agtattgcaa ctactggcgt aagcttatag ttggtaccac
     2701 attaccacat taccacttgt taatgtgtgt tactttcatg agaaatgcta aaatataaaa
     2761 gccagagtgt ggcagcactc tagcttttaa aaaagaaact accctttaat aggagggtcc
     2821 tcgtgtagac acttttcctt tacagtaatg taaataggat gagtgtcttt ttttctaagc
     2881 atgaataata atgttaaaag agatattacc catgtcataa tcccgcctat taaattggca
     2941 aagccgatca tgaccattag agattcatag gtcgacatag tagttcctcc ttatgtaagc
     3001 ttaattgtta tccgctcaca attacacaca ttatgccaca ccttgtagat aaagtcaaca
     3061 acttttgcaa aatgaattgt gagtgctcac atttaccctc gagcaacgtt cttgccattg
     3121 ctgcataaaa aacgcccggc ggcaaccgag cgttctgaat taattaatca tcgggaagat
     3181 cttcatcacc gaaacgcggc aggcagctct agagttaaca agagtttgta gaaacgcaaa
     3241 aaggccatcc gtcaggatgg ccttctgctt agctagagcg gcggatttgt cctactcagg
     3301 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     3361 cgttttattt gatgcctcaa gctagaggtc gaattcctgc agccctggcc gagaccggtc
     3421 tcggaatggc gattttcgtt cgtgaataca tgttataata actataacta ataacgtaac
     3481 gtgactggca agagatattt ttaaaacaat gaataggttt acacttactt tagttttatg
     3541 gaaatgaaag atcatatcat atataatcta gaataaaatt aactaaaata attattatct
     3601 agataaaaaa tttagaagcc aatgaaatct ataaataaac taaattaagt ttatttaatt
     3661 aacaactatg gatataaaat aggtactaat caaaatagtg aggaggatat atttgaatac
     3721 atacgaacaa attaataaag tgaaaaaaat acttcggaaa catttaaaaa ataaccttat
     3781 tggtacttac atgtttggat caggagttga gagtggacta aaaccaaata gtgatcttga
     3841 ctttttagtc gtcgtatctg aaccattgac agatcaaagt aaagaaatac ttatacaaaa
     3901 aattagacct atttcaaaaa aaataggaga taaaagcaac ttacgatata ttgaattaac
     3961 aattattatt cagcaagaaa tggtaccgtg gaatcatcct cccaaacaag aatttattta
     4021 tggagaatgg ttacaagagc tttatgaaca aggatacatt cctcagaagg aattaaattc
     4081 agatttaacc ataatgcttt accaagcaaa acgaaaaaat aaaagaatat acggaaatta
     4141 tgacttagag gaattactac ctgatattcc attttctgat gtgagaagag ccattatgga
     4201 ttcgtcagag gaattaatag ataattatca ggatgatgaa accaactcta tattaacttt
     4261 atgccgtatg attttaacta tggacacggg taaaatcata ccaaaagata ttgcgggaaa
     4321 tgcagtggct gaatcttctc cattagaaca tagggagaga attttgttag cagttcgtag
     4381 ttatcttgga gagaatattg aatggactaa tgaaaatgta aatttaacta taaactattt
     4441 aaataacaga ttaaaaaaat tataaaaaaa ttgaaaaaat ggtggaaaca cttttttcaa
     4501 tttttttgtt ttattattta atatttggga aatattcatt ctaattggta atcagatttt
     4561 agaaaacaat aaacccttgc atagggggaa tccgtttagg ctgggcggtg atagcttctc
     4621 gttcaggcag tacgcctctt ttcttttcca gacctgaggg aggcggaaat ggtgtgaggt
     4681 tcccggggaa aagccaaata ggcgatcgcg ggagtgcttt atttgaagat caggctatca
     4741 ctgcggtcaa tagatttcac aatgtgatgg ctggacagcc tgaggaactc tcgaacccga
     4801 atggaaacaa ccagatattt atgaatcagc gcggctcaca tggcgttgtg ctggcaaatg
     4861 caggttcatc ctctgtctct atcaatacgg caacaaaatt gcctgatggc aggtatgaca
     4921 ataaagctgg agcgggttca tttcaagtga acgatggtaa actgacaggc acgatcaatg
     4981 ccaggtctgt agctgtgctt tatcctgatg atattgcaaa agcgcctcat gttttccttg
     5041 agaattacaa aacaggtgta acacattctt tcaatgatca actgacgatt accttgcgtg
     5101 cagatgcgaa tacaacaaaa gccgtttatc aaatcaataa tggaccagag acggcgttta
     5161 aggatggaga tcaattcaca atcggaaaag gagatccatt tggcaaaaca tacaccatca
     5221 tgttaaaagg aacgaacagt gatggtgtaa cgaggaccga gaaatacagt tttgttaaaa
     5281 gagatccagc gtcggccaaa accatcggct atcaaaatcc gaatcattgg agccaggtaa
     5341 atgcttatat ctataaacat gatgggagcc gagtaattga attgaccgga tcttggcctg
     5401 gaaaaccaat gactaaaaat gcagacggaa tttacacgct gacgctgcct gcggacacgg
     5461 atacaaccaa cgcaaaagtg atttttaata atggcagcgc ccaagtgccc ggtcagaatc
     5521 agcctggctt tgattacgtg ctaaatggtt tatataatga ctcgggctta agcggttctc
     5581 ttccccattg agggcaaggc tagacgggac ttaccgaaag aaaccatcaa tgatggtttc
     5641 ttttttgttc ataaatcaga caaaactttt ctcttgcaaa agtttgtgaa gtgttgcaca
     5701 atataaatgt gaaatacttc acaaacaaaa agacatcaaa gagaaacata ccctgcaagg
     5761 atgctgatat tgtctgcatt tgcgccggag caaaccaaaa acctggtgag acacgccttg
     5821 aattagtaga aaagaacttg aagattttca aaggcatcgt tagtgaagtc atggcgagcg
     5881 gatttgacgg cattttctta gtcgcgacgc gaggctggat ggccttcccc attatgattc
     5941 ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag
     6001 atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga
     6061 tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt
     6121 tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     6181 catggagccg ggccacctac tgaagtggat ttctttaaga gctcctttaa cttcctcacc
     6241 agtagttgta tcggtaccat aagtagaagc agcaacccaa gtagctttac cagcatccgg
     6301 ttcaaccagc atagtaagaa tcttactgga catcggcagt tcttcgaaca gtgcgccaac
     6361 taccagctct ttctgcagtt cattcagggc accggagaac ctgcgtgcaa tccatcttgt
     6421 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcca tggattacgc gttaacccgg
     6481 gcccgcggat gcatatgatc agatcttaag gcctaggtct agagtctttg ttttgacgcc
     6541 attagcgtac gtaacaatcc tcgttaaagg acaaggacct gagcggaagt gtatcgtaca
     6601 gtagacggag tatactagta tagtctatag tccgtggaat tattatattt atctccgacg
     6661 atattctcat cagtgaaatc cagctggagt tctttagcaa atttttttat tagctgaact
     6721 tagtattagt ggccatactc ctccaatcca aagctattta gaaagattac tatatcctca
     6781 aacaggcggt aaccggcctc ttcatcggga atgcgcgcga ccttcagcat cgccggcatg
     6841 tccccctggc ggacgggaag tatccagctc gaggtcgggc cgcgttgctg gcgtttttcc
     6901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7321 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7501 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7561 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc
     7801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     7981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     8281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8641 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     8701 cgaggccctt tcgtcttcaa gaatt
//
