LOCUS       sequence-446181-        5812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive GFP expression for Bacillus subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5812)
  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 5812)
  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: pOSV00170
FEATURES             Location/Qualifiers
     source          1..5812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..144
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(328..375)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    2117..2133
                     /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)."
     primer_bind     complement(3563..3582)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3564..3582)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3667..3684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3838..4426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3918..3937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4597..5388,5389..5457))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5220..5239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5458..5562)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5588..5812,1..231))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5725..5746)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
       61 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
      121 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
      181 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
      241 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      301 agatccggat atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc
      361 aaggggttat gctagttatt gctcagccgg ccgctcagac tgttttcggg aattgtcatt
      421 ttagtctttt tcacgtttta tgtatcgtcc ggaatggtgt ccggcggcgt cttattcaac
      481 agcatcttgg ggatggatta tcacaccggc ctgtggattg tgacgggtgt tgttgtggca
      541 tataccctgt ttggcggctt tttggcagtc agctggacag attttgttca ggggattatt
      601 atgtttgccg cactcattct tgtgccgatc gttacgtttt tccacacggg aggagcaggt
      661 gatacagttg ctgaaatccg ctctgtagat cctgatatgt ttaatatttt caaaggaacg
      721 agcgtccttg gcattatttc tttgtttgcc tgggggctgg gatactttgg ccagccgcat
      781 attattgtgc gttttatggc gataacgtct gtcaaagaga tcaaaagagc gcgcagaatt
      841 gggatgggct ggatgatttt gtctgcagta ggcgcagtgc tgaccggttt gggcggaatc
      901 gcttattacc atcagcgcgg catgacactg aaggacccag agacgatttt tattcaatta
      961 gggaatatct tgttccatcc gattatcacg gggtcgacgg agctgtaata taaaaacctt
     1021 cttcaactaa cggggcaggt tagtgacatt agaaaaccga ctgtaaaaag tacagtcggc
     1081 attatctcat attataaaag ccagtcatta ggcctatctg acaattcctg aatagagttc
     1141 ataaacaatc ctgcatgata accatcacaa acagaatgat gtacctgtaa agatagcggt
     1201 aaatatattg aattaccttt attaatgaat tttcctgctg taataatggg tagaaggtaa
     1261 ttactattat tattgatatt taagttaaac ccagtaaatg aagtccatgg aataatagaa
     1321 agagaaaaag cattttcagg tataggtgtt ttgggaaaca atttccccga accattatat
     1381 ttctctacat cagaaaggta taaatcataa aactctttga agtcattctt tacaggagtc
     1441 caaataccag agaatgtttt agatacacca tcaaaaattg tataaagtgg ctctaactta
     1501 tcccaataac ctaactctcc gtcgctattg taaccagttc taaaagctgt atttgagttt
     1561 atcacccttg tcactaagaa aataaatgca gggtaaaatt tatatccttc ttgttttatg
     1621 tttcggtata aaacactaat atcaatttct gtggttatac taaaagtcgt ttgttggttc
     1681 aaataatgat taaatatctc ttttctcttc caattgtcta aatcaatttt attaaagttc
     1741 atttgatatg cctcctaaat ttttatctaa agtgaattta ggaggcttac ttgtctgctt
     1801 tcttcattag aatcaatcct tttttaaaag tcaatattac tgtaacataa atatatattt
     1861 taaaaatatc ccactttatc caattttcgt ttgttgaact aatgggtgct ttagttgaag
     1921 aataaaagac cacattaaaa aatgtggtct tttgtgtttt tttaaaggat ttgagcgtag
     1981 cgaaaaatcc ttttctttct tatcttgata ataagggtaa ctattgccgt atgggatccg
     2041 ggtaaatgtg agcactcaca attcattttg caaaagttgt tgactttatc tacaaggtgt
     2101 ggcataatgt gtgtaattgt gagcggataa caattaagct tagtcgacag ctagctgatt
     2161 aactaataag gaggacaaac atggtttcta aaggtgaaga attgtttaca ggtgttgttc
     2221 caattttggt tgaattggat ggtgatgtta atggtcataa attttctgtt tctggtgaag
     2281 gtgaaggtga tgctacatac ggtaaattga cattgaaatt tatttgtaca actggtaaat
     2341 tgccagttcc ttggccaaca ttggttacaa catttgctta tggtttgcaa tgttttgctc
     2401 gttatccaga tcacatgaaa caacatgatt tctttaaatc tgctatgcca gaaggttatg
     2461 ttcaagaacg tacaatcttt ttcaaggatg atggtaatta taagacacgt gctgaggtta
     2521 agtttgaagg tgatacattg gttaatcgta tcgaattgaa gggtatcgat tttaaagaag
     2581 atggtaatat cttgggtcat aaattggaat ataattataa ttctcataat gtttatatca
     2641 tggctgataa acaaaagaac ggtattaaag ttaattttaa aattcgtcat aatattgaag
     2701 atggttctgt tcaattggct gatcattatc aacaaaatac accaattggt gatggtccag
     2761 ttttgttgcc agataatcat tatttgtcta cacaatctaa attgtctaaa gatccaaatg
     2821 aaaaacgtga tcacatggtt ttgttggaat ttgttacagc tgctggtatt acacatggta
     2881 tggatgaatt gtataaataa gaattccttc ccagctgctt gtgacatcaa gctctctggt
     2941 agaagatttg tataagtcga tgttcagacg ctcagcttca gataaagaac tggtgttttt
     3001 aggccgactg gctgttctgg ctgtttcaat tgtcgcgttg gttctggctt gggagaaaaa
     3061 taacaccatc cttggattgg tcagctacgc gtgggcgggc ttcggtgcat cattcgggcc
     3121 agttgtactg ctcagtctgt tttggaaacg aatgaccaaa tggggggcac tcgccggcat
     3181 gattgtggga gcagcgactg taatcatttg ggcgaatgcc ggtctttcgg actttctgta
     3241 tgaaatgata cctggttttg ctgcgagtct attatctgtc ttttttgtca gcatattgac
     3301 gcaggctccg tcacaagctg tcacagacca gttcaacgac taccaagata caatgtcgca
     3361 ataaagatcg aaaggaggag gaaggggctg tccctttttc ctcttttctc cttttaagcg
     3421 gccatttttt tcttttggaa acaaacgtat ttttttctat acaaaaagac cccgtatgga
     3481 tatgattaaa gtaaaaaaag tataggagaa aaaggtatgg aagagctttt agagagagtc
     3541 tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg ggatcgagat
     3601 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3661 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4021 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4261 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5581 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5641 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5701 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5761 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact ac
//
