LOCUS       sequence-252686-        8814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible system (gfpmut2 with Ppsank and riboJ) for expression
            in B. subtilis .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8814)
  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 8814)
  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: pLG231
FEATURES             Location/Qualifiers
     source          1..8814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      261..347
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      366..393
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      473..507
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    635..651
                     /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)."
     misc_RNA        656..705
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             753..1469
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(780..808)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1403..1423
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1519..1613
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1819..2901
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1838..1857)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3754..3772)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3840..3944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3945..4013,4014..4805)
                     /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(4163..4182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4976..5564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5465..5484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatttagtt tatttataga tttcattggc ttctaaattt tttatctaga taataattat
       61 tttagttaat tttattctag attatatatg atatgatctt tcatttccat aaaactaaag
      121 taagtgtaaa cctattcatt gttttaaaaa tatctcttgc cagtcacgtt acgttattag
      181 ttatagttat tataacatgt attcacgaac gaaaatcgcc attcgccagc tgcaggaatt
      241 cgacctctag cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      301 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgctctagc
      361 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttgttaactc
      421 tagagctgcc tgccgcgttt cggtgatgaa gatcttcccg atgattaatt aattcagaac
      481 gctcggttgc cgccgggcgt tttttatgca gcaatggcaa gaacgttgct cgagggtctc
      541 ccgcttaacg atcgttggct gaaatgtgag cactcacaat tcattttgca aaagttgttg
      601 actttatcta caaggtgtgg cataatgtgt gtaattgtga gcggataaca attagctgtc
      661 accggatgtg ctttccggtc tgatgagtcc gtgaggacga aacagcctct acaaataatt
      721 ttgtttaatc ggctgaaagg tggtgaacta ctatgaaagg agaagaactt ttcactggag
      781 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
      841 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
      901 gaaaactacc tgttccatgg ccaacacttg tcactacttt cgcgtatggt cttcaatgct
      961 ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     1021 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     1081 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     1141 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     1201 acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca
     1261 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     1321 gccctgtcct tttaccagac aaccattacc tgtccacaca atctaagctt tcgaaagatc
     1381 ccaacgaaaa gagggaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     1441 atggcatgga tgaactatac aaatactagt aagcttgcgg ccgcgtcgtg actgggaaaa
     1501 ccctggcgac tagtcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1561 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccagggg
     1621 tccccaataa ttacgattta aattggagct tagtcgacag ctagccgcat gcaagctaat
     1681 tcggtggaaa cgaggtcatc atttccttcc gaaaaaacgg ttgcatttaa atcttacata
     1741 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgtagcgccc ggaagagagt
     1801 caattcaggg tggtgaatat gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     1861 gtctcttata tgaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     1921 cgggaaaaag tggaagcggc gatggtggag ctgaattaca ttcccaaccg cgtggcacaa
     1981 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     2041 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     2101 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     2161 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     2221 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     2281 cccatcaaca gtattattta ctcccatgag gacggtacgc gactgggcgt ggagcatctg
     2341 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     2401 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     2461 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     2521 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     2581 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     2641 gataccgaag atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     2701 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     2761 ggcaatcagc tgttgccagt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     2821 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2881 cgactggaaa gcgggcagtg ataaggatcc taattggtaa cgaatcagac aattgacggc
     2941 gcgcaacgca attaatgtga gttaggatcc tgagcgccgg tcgctaccat taccagttgg
     3001 tctggtgtca aaaataataa taaccgggca ggccatgtct gcccgtattt cgcgtaagga
     3061 aatccattat gtactattta attctgcgtg acatcccatc gatcagacca gtttttaatt
     3121 tgtgtgtttc catgtgtcca gtttggaata ctcttaacct cattggaaat cgcggcataa
     3181 tcactggtgg tatgattgat gaccgcgtca acaatgacct ttatgccata ttcttcagcg
     3241 gctgcacaca tttctttaaa ttcttgttca gtacctaagt aacggttgcc aatttgatac
     3301 gatgtcggct gatacagcca gtaccagttc gacatgcttt tatctccttg attcccttcc
     3361 tttacttggt taatcggaga tgtctgaatg gctgtatatc ctgcatcatg aatatccttc
     3421 atattgtgtt ttaacgtatt gaacgaccaa ttccatgcat gaagaatggt tccgcttttg
     3481 atcgacggtg ctgtaagctc attcgatttg ttcgccgttt cagcactcgc agccgccggt
     3541 cctgccagaa ccaaatgaaa cagcaataaa aatccagcga ataacggcag taaagaggtt
     3601 ttgaatcgtt ttgcaaacat tcttgacact ccttatttga ttttttgaag acttacttcg
     3661 gagtcaaaaa tccctcttac ttcattcttc cgcttcctcc tttcaaaccg atgtgaagac
     3721 tggagaattt tgttaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     3781 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3841 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3901 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3961 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4021 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4081 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4141 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     4201 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4261 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4321 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4381 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4441 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4501 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4561 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4621 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtcgcgcggt atcattgcag
     4681 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4741 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4801 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4861 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4921 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4981 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5041 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5101 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5161 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5221 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5281 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5341 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5401 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5461 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5521 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5581 cccgacctcg agctggatac ttcccgtccg ccagggggac atgccggcga tgctgaaggt
     5641 cgcgcgcatt cccgatgaag aggccggtta ccgcctgttt gaggatatag taatctttct
     5701 aaatagcttt ggattggagg agtatggcca ctaatactaa gttcagctaa taaaaaaatt
     5761 tgctaaagaa ctccagctgg atttcactga tgagaatatc gtcggagata aatataataa
     5821 ttccacggac tatagactat actagtatac tccgtctact gtacgataca cttccgctca
     5881 ggtccttgtc ctttaacgag gattgttacg tacgctaatg gcgtcaaaac aaagactcta
     5941 gacctaggcc ttaagatctg atcatatgca tccgcgggcc cgggttaacg cgtaatccat
     6001 ggatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     6061 ttctccggtg ccctgaatga actgcagaaa gagctggtag ttggcgcact gttcgaagaa
     6121 ctgccgatgt ccagtaagat tcttactatg ctggttgaac cggatgctgg taaagctact
     6181 tgggttgctg cttctactta tggtaccgat acaactactg gtgaggaagt taaaggagct
     6241 cttaaagaaa tccacttcag taggtggccc ggctccatgc accgcgacgc aacgcgggga
     6301 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     6361 aggcggtata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     6421 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     6481 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     6541 tccagcctcg cgtcgcgact aagaaaatgc cgtcaaatcc gctcgccatg acttcactaa
     6601 cgatgccttt gaaaatcttc aagttctttt ctactaattc aaggcgtgtc tcaccaggtt
     6661 tttggtttgc tccggcgcaa atgcagacaa tatcagcatc cttgcagggt atgtttctct
     6721 ttgatgtctt tttgtttgtg aagtatttca catttatatt gtgcaacact tcacaaactt
     6781 ttgcaagaga aaagttttgt ctgatttatg aacaaaaaag aaaccatcat tgatggtttc
     6841 tttcggtaag tcccgtctag ccttgccctc aatggggaag agaaccgctt aagcccgagt
     6901 cattatataa accatttagc acgtaatcaa agccaggctg attctgaccg ggcacttggg
     6961 cgctgccatt attaaaaatc acttttgcgt tggttgtatc cgtgtccgca ggcagcgtca
     7021 gcgtgtaaat tccgtctgca tttttagtca ttggttttcc aggccaagat ccggtcaatt
     7081 caattactcg gctcccatca tgtttataga tataagcatt tacctggctc caatgattcg
     7141 gattttgata gccgatggtt ttggccgacg ctggatctct tttaacaaaa ctgtatttct
     7201 cggtcctcgt tacaccatca ctgttcgttc cttttaacat gatggtgtat gttttgccaa
     7261 atggatctcc ttttccgatt gtgaattgat ctccatcctt aaacgcctgt cgtctggtcc
     7321 attattgatt tgataaacgg cttttgttgt attcgcatct gcacgcaagg taatcgtcag
     7381 ttgatcattg aaagaatgtg ttacacctgt tttgtaattc tcaaggaaaa catgaggcgc
     7441 ttttgcaata tcatcaggat aaagcacagc tacagacctg gcattgatcg tgcctgtcag
     7501 tttaccatcg ttcacttgaa atgaacccgc tccagcttta ttgtcatacc tgccatcagg
     7561 caattttgtt gccgtattga tagagacaga ggatgaacct gcatttgcca gcacaacgcc
     7621 atgtgagccg cgctgattca taaatatctg gttgtttcca ttcgggttcg agagttcctc
     7681 aggctgtcca gccatcacat tgtgaaatct attgaccgca gtgatagcct gatcttcaaa
     7741 taaagcactc ccgcgatcgc ctatttggct tttccccggg aacctcacac catttccgcc
     7801 tccctcaggt ctggaaaaga aaagaggcgt actgcctgaa cgagaagcta tcaccgccca
     7861 gcctaaacgg attcccccta tgcaagggtt tattgttttc taaaatctga ttaccaatta
     7921 gaatgaatat ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa gtgtttccac
     7981 cattttttca atttttttat aattttttta atctgttatt taaatagttt atagttaaat
     8041 ttacattttc attagtccat tcaatattct ctccaagata actacgaact gctaacaaaa
     8101 ttctctccct atgttctaat ggagaagatt cagccactgc atttcccgca atatcttttg
     8161 gtatgatttt acccgtgtcc atagttaaaa tcatacggca taaagttaat atagagttgg
     8221 tttcatcatc ctgataatta tctattaatt cctctgacga atccataatg gctcttctca
     8281 catcagaaaa tggaatatca ggtagtaatt cctctaagtc ataatttccg tatattcttt
     8341 tattttttcg ttttgcttgg taaagcatta tggttaaatc tgaatttaat tccttctgag
     8401 gaatgtatcc ttgttcataa agctcttgta accattctcc ataaataaat tcttgtttgg
     8461 gaggatgatt ccacggtacc atttcttgct gaataataat tgttaattca atatatcgta
     8521 agttgctttt atctcctatt ttttttgaaa taggtctaat tttttgtata agtatttctt
     8581 tactttgatc tgtcaatggt tcagatacga cgactaaaaa gtcaagatca ctatttggtt
     8641 ttagtccact ctcaactcct gatccaaaca tgtaagtacc aataaggtta ttttttaaat
     8701 gtttccgaag tatttttttc actttattaa tttgttcgta tgtattcaaa tatatcctcc
     8761 tcactatttt gattagtacc tattttatat ccatagttgt taattaaata aact
//
