LOCUS       sequence-252556-        8382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vanillic acid inducible system for expression in B. subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8382)
  AUTHORS   Gonzalez LM, Mukhitov N, Voigt CA
  TITLE     Resilient living materials built by printing bacterial spores.
  JOURNAL   Nat Chem Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5.
  PUBMED    31792444
REFERENCE   2  (bases 1 to 8382)
  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 Chem 
            Biol. 2019 Dec 2. pii: 10.1038/s41589-019-0412-5. doi: 
            10.1038/s41589-019-0412-5."
COMMENT     Sequence Label: pLG320
FEATURES             Location/Qualifiers
     source          1..8382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2586..2672
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2691..2718
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        2905..2954
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             3002..3718
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(3029..3057)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3652..3672
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3768..3862
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(5652..5670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5738..5842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5843..5911,5912..6703)
                     /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     complement(6061..6080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6874..7462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7363..7382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
       61 cgcgtcgcga ctaagaaaat gccgtcaaat ccgctcgcca tgacttcact aacgatgcct
      121 ttgaaaatct tcaagttctt ttctactaat tcaaggcgtg tctcaccagg tttttggttt
      181 gctccggcgc aaatgcagac aatatcagca tccttgcagg gtatgtttct ctttgatgtc
      241 tttttgtttg tgaagtattt cacatttata ttgtgcaaca cttcacaaac ttttgcaaga
      301 gaaaagtttt gtctgattta tgaacaaaaa agaaaccatc attgatggtt tctttcggta
      361 agtcccgtct agccttgccc tcaatgggga agagaaccgc ttaagcccga gtcattatat
      421 aaaccattta gcacgtaatc aaagccaggc tgattctgac cgggcacttg ggcgctgcca
      481 ttattaaaaa tcacttttgc gttggttgta tccgtgtccg caggcagcgt cagcgtgtaa
      541 attccgtctg catttttagt cattggtttt ccaggccaag atccggtcaa ttcaattact
      601 cggctcccat catgtttata gatataagca tttcctggct ccaatgattc ggattttgat
      661 agccgatggt tttggccgac gctggatctc ttttaacaaa actgtatttc tcggtcctcg
      721 ttacaccatc actgttcgtt ccttttaaca tgatggtgta tgttttgcca aatggatctc
      781 cttttccgat tgtgaattga tctccatcct taaacgcctg tcgtctggtc cattattgat
      841 ttgataaacg gcttttgttg tattcgcatc tgcacgcaag gtaatcgtca gttgatcatt
      901 gaaagaatgt gttacacctg ttttgtaatt ctcaaggaaa acatgaggcg cttttgcaat
      961 atcatcagga taaagcacag ctacagacct ggcattgatc gtgcctgtca gtttaccatc
     1021 gttcacttga aatgaacccg ctccagcttt attgtcatac ctgccatcag gcaattttgt
     1081 tgccgtattg atagagacag aggatgaacc tgcatttgcc agcacaacgc catgtgagcc
     1141 gcgctgattc ataaatatct ggttgtttcc attcgggttc gagagttcct caggctgtcc
     1201 agccatcaca ttgtgaaatc tattgaccgc agtgatagcc tgatcttcaa ataaagcact
     1261 cccgcgatcg cctatttggc ttttccccgg gaacctcaca ccatttccgc ctccctcagg
     1321 tctggaaaag aaaagaggcg tactgcctga acgagaagct atcaccgccc agcctaaacg
     1381 gattccccct atgcaagggt ttattgtttt ctaaaatctg attaccaatt agaatgaata
     1441 tttcccaaat attaaataat aaaacaaaaa aattgaaaaa agtgtttcca ccattttttc
     1501 aattttttta taattttttt aatctgttat ttaaatagtt tatagttaaa tttacatttt
     1561 cattagtcca ttcaatattc tctccaagat aactacgaac tgctaacaaa attctctccc
     1621 tatgttctaa tggagaagat tcagccactg catttcccgc aatatctttt ggtatgattt
     1681 tacccgtgtc catagttaaa atcatacggc ataaagttaa tatagagttg gtttcatcat
     1741 cctgataatt atctattaat tcctctgacg aatccataat ggctcttctc acatcagaaa
     1801 atggaatatc aggtagtaat tcctctaagt cataatttcc gtatattctt ttattttttc
     1861 gttttgcttg gtaaagcatt atggttaaat ctgaatttaa ttccttctga ggaatgtatc
     1921 cttgttcata aagctcttgt aaccattctc cataaataaa ttcttgtttg ggaggatgat
     1981 tccacggtac catttcttgc tgaataataa ttgttaattc aatatatcgt aagttgcttt
     2041 tatctcctat tttttttgaa ataggtctaa ttttttgtat aagtatttct ttactttgat
     2101 ctgtcaatgg ttcagatacg acgactaaaa agtcaagatc actatttggt tttagtccac
     2161 tctcaactcc tgatccaaac atgtaagtac caataaggtt attttttaaa tgtttccgaa
     2221 gtattttttt cactttatta atttgttcgt atgtattcaa atatatcctc ctcactattt
     2281 tgattagtac ctattttata tccatagttg ttaattaaat aaacttaatt tagtttattt
     2341 atagatttca ttggcttcta aattttttat ctagataata attattttag ttaattttat
     2401 tctagattat atatgatatg atctttcatt tccataaaac taaagtaagt gtaaacctat
     2461 tcattgtttt aaaaatatct cttgccagtc acgttacgtt attagttata gttattataa
     2521 catgtattca cgaacgaaaa tcgccattcg ccagctgcag gaattcgacc tctagcttga
     2581 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2641 tgtcggtgaa cgctctcctg agtaggacaa atccgccgct ctagctaagc agaaggccat
     2701 cctgacggat ggcctttttg cgtttctaca aactcttgtt aactctagag ctgcctgccg
     2761 cgtttcggtg atgaagatct tcccgatgat tattaattca gaacgctcgg ttgccgcggc
     2821 aagaacgttg ctcgagattg gatccaatca ttttgcaaaa gttgttgact ttatctacaa
     2881 ggtgtggcat attggatcca atagctgtca ccggatgtgc tttccggtct gatgagtccg
     2941 tgaggacgaa acagcctcta caaataattt tgtttaatcg gctgaaaggt ggtgaactac
     3001 tatgaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     3061 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     3121 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt
     3181 cactactttc gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca
     3241 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
     3301 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
     3361 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt
     3421 ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat
     3481 caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     3541 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     3601 gtccacacaa tctaagcttt cgaaagatcc caacgaaaag agggaccaca tggtccttct
     3661 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aatactagta
     3721 agcttgcggc cgcgtcgtga ctgggaaaac cctggcgact agtcttggac tcctgttgat
     3781 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     3841 ggcgtttttt attggtgaga atccaggggt ccccaataat tacgatttaa attggagctt
     3901 agtcgacagc tagccgcatg caagctaatt cggtggaaac gaggtcatca tttccttccg
     3961 aaaaaacggt tgcatttaaa tcttacatat gtaatacttt caaagactac atttgtaaga
     4021 tttgatgttt gtagcgcccg tcggctgaaa ggtggtgaac tactatggac atgcctcgta
     4081 ttaaaccggg tcagcgtgtt atgatggcac tgcgtaaaat gattgcaagc ggtgaaatca
     4141 aaagtggtga acgtattgca gaaattccga ccgcagcagc actgggtgtt agccgtatgc
     4201 cggttcgtat cgcactgcgt tcactggaac aagaaggtct ggttgttcgt ctgggtgcac
     4261 gtggttatgc agcccgtggt gttagcagcg atcagattcg tgatgcaatt gaagttcgtg
     4321 gtgttctgga aggttttgca gcacgtcgtc tggcagaacg tggtatgacc gcagaaaccc
     4381 atgcacgttt tgttgtactg attgcagaag gtgaagcact gtttgcagcc ggtcgcctga
     4441 atggtgaaga tctggatcgt tatgccgcat ataatcaggc atttcatgat accctggtta
     4501 gcgcagcagg taatggtgca gttgaaagcg cactggcacg taatggtttt gaaccgtttg
     4561 cagcagccgg tgcactggcc ctggatctga tggacctgtc tgccgaatat gaacatctgc
     4621 tggcagcaca tcgtcagcat caggcagttc tggatgcagt tagctgtggt gatgccgaag
     4681 gtgcagaacg tattatgcgt gatcatgcac tggcagcaat tcgtaatgca aaagtttttg
     4741 aagcagcagc aagcgcaggc gcaccgctgg gtgcagcatg gtcaattcgt gcagattgat
     4801 aaggatccta attggtaacg aatcagacaa ttgacggcgc gcaacgcaat taatgtgagt
     4861 taggatcctg agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataataata
     4921 accgggcagg ccatgtctgc ccgtatttcg cgtaaggaaa tccattatgt actatttaat
     4981 tctgcgtgac atcccatcga tcagaccagt ttttaatttg tgtgtttcca tgtgtccagt
     5041 ttggaatact cttaacctca ttggaaatcg cggcataatc actggtggta tgattgatga
     5101 ccgcgtcaac aatgaccttt atgccatatt cttcagcggc tgcacacatt tctttaaatt
     5161 cttgttcagt acctaagtaa cggttgccaa tttgatacga tgtcggctga tacagccagt
     5221 accagttcga catgctttta tctccttgat tcccttcctt tacttggtta atcggagatg
     5281 tctgaatggc tgtatatcct gcatcatgaa tatccttcat attgtgtttt aacgtattga
     5341 acgaccaatt ccatgcatga agaatggttc cgcttttgat cgacggtgct gtaagctcat
     5401 tcgatttgtt cgccgtttca gcactcgcag ccgccggtcc tgccagaacc aaatgaaaca
     5461 gcaataaaaa tccagcgaat aacggcagta aagaggtttt gaatcgtttt gcaaacattc
     5521 ttgacactcc ttatttgatt ttttgaagac ttacttcgga gtcaaaaatc cctcttactt
     5581 cattcttccg cttcctcctt tcaaaccgat gtgaagactg gagaattttg ttaattcttg
     5641 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5701 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6061 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     6121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6361 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     6421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6541 tggagccggt gagcgtgggt cgcgcggtat cattgcagca ctggggccag atggtaagcc
     6601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7021 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7381 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7441 aggggggcgg agcctatgga aaaacgccag caacgcggcc cgacctcgag ctggatactt
     7501 cccgtccgcc agggggacat gccggcgatg ctgaaggtcg cgcgcattcc cgatgaagag
     7561 gccggttacc gcctgtttga ggatatagta atctttctaa atagctttgg attggaggag
     7621 tatggccact aatactaagt tcagctaata aaaaaatttg ctaaagaact ccagctggat
     7681 ttcactgatg agaatatcgt cggagataaa tataataatt ccactataga ctatactagt
     7741 atactccgtc tactgtacga tacacttccg ctcaggtcct tgtcctttaa cgaggattgt
     7801 tacgtacgct aatggcgtca aaacaaagac tctagaccta ggccttaaga tctgatcata
     7861 tgcatccgcg ggcccgggtt aacgcgtaat ccatggatca agagacagga tgaggatcgt
     7921 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggtgccctga atgaactgca
     7981 gaaagagctg gtagttggcg cactgttcga agaactgccg atgtccagta agattcttac
     8041 tatgctggtt gaaccggatg ctggtaaagc tacttgggtt gctgcttcta cttatggtac
     8101 cgatacaact actggtgagg aagttaaagg agctcttaaa gaaatccact tcagtaggtg
     8161 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     8221 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg tataaatcgc cgtgacgatc
     8281 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     8341 tgatggtcgt catctacctg cctggacagc atggcctgca ac
//
