LOCUS       sequence-252599-        8385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xylose inducible system without CRE site; cell do not need minimal 
            media to express gfp under this modify xylose promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8385)
  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 8385)
  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: pLG327
FEATURES             Location/Qualifiers
     source          1..8385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      120..191
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        278..327
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             375..1091
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(402..430)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1025..1045
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1120..1167
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1884..1902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1970..2074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2075..2143,2144..2935)
                     /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(2293..2312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3106..3694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3595..3614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtcatttca gataattttg ctcttgaaat aggtgaattc ttaagtattt cttttaataa
       61 taacttttga tttacttttt tgacaaaggt ttgatctgcg atatccactt ctgaggcatc
      121 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
      181 tgaacgctct catgaaaaac taaaaaaaat attgaaaata ctgatgaggt tatttaagat
      241 taaaataagt tagtttgttt gggcaacaaa ctaatagctg tcaccggatg tgctttccgg
      301 tctgatgagt ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa tcggctgaaa
      361 ggtggtgaac tactatgaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      421 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      481 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
      541 ggccaacact tgtcactact ttcgcgtatg gtcttcaatg ctttgcgaga tacccagatc
      601 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
      661 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
      721 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
      781 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
      841 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
      901 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
      961 acaaccatta cctgtccaca caatctaagc tttcgaaaga tcccaacgaa aagagggacc
     1021 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     1081 acaaatacta gctgcattca gcccgcctaa tgagcgggcc tagcataacc ccttggggcc
     1141 tctaaacggg tcttgagggg ttttttggga gcccgtattt cgcgtaagga aatccattat
     1201 gtactattta attctgcgtg acatcccatc gatcagacca gtttttaatt tgtgtgtttc
     1261 catgtgtcca gtttggaata ctcttaacct cattggaaat cgcggcataa tcactggtgg
     1321 tatgattgat gaccgcgtca acaatgacct ttatgccata ttcttcagcg gctgcacaca
     1381 tttctttaaa ttcttgttca gtacctaagt aacggttgcc aatttgatac gatgtcggct
     1441 gatacagcca gtaccagttc gacatgcttt tatctccttg attcccttcc tttacttggt
     1501 taatcggaga tgtctgaatg gctgtatatc ctgcatcatg aatatccttc atattgtgtt
     1561 ttaacgtatt gaacgaccaa ttccatgcat gaagaatggt tccgcttttg atcgacggtg
     1621 ctgtaagctc attcgatttg ttcgccgttt cagcactcgc agccgccggt cctgccagaa
     1681 ccaaatgaaa cagcaataaa aatccagcga ataacggcag taaagaggtt ttgaatcgtt
     1741 ttgcaaacat tcttgacact ccttatttga ttttttgaag acttacttcg gagtcaaaaa
     1801 tccctcttac ttcattcttc cgcttcctcc tttcaaaccg atgtgaagac tggagaattt
     1861 tgttaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1921 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccatg
     2041 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2161 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtt ggttacatcg aactggatct
     2221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2281 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2341 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2461 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2521 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2581 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     2641 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2701 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2761 tgctgataaa tctggagccg gtgagcgtgg gtcgcgcggt atcattgcag cactggggcc
     2821 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2881 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2941 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3001 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cccgacctcg
     3721 agctggatac ttcccgtccg ccagggggac atgccggcga tgctgaaggt cgcgcgcatt
     3781 cccgatgaag aggccggtta ccgcctgttt gaggatatag taatctttct aaatagcttt
     3841 ggattggagg agtatggcca ctaatactaa gttcagctaa taaaaaaatt tgctaaagaa
     3901 ctccagctgg atttcactga tgagaatatc gtcggagata aatataataa ttccacggac
     3961 tatagactat actagtatac tccgtctact gtacgataca cttccgctca ggtccttgtc
     4021 ctttaacgag gattgttacg tacgctaatg gcgtcaaaac aaagactcta gacctaggcc
     4081 ttaagatctg atcatatgca tccgcgggcc cgggttaacg cgtaatccat ggatcaagag
     4141 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggtg
     4201 ccctgaatga actgcagaaa gagctggtag ttggcgcact gttcgaagaa ctgccgatgt
     4261 ccagtaagat tcttactatg ctggttgaac cggatgctgg taaagctact tgggttgctg
     4321 cttctactta tggtaccgat acaactactg gtgaggaagt taaaggagct cttaaagaaa
     4381 tccacttcag taggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     4441 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata
     4501 aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga
     4561 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     4621 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     4681 cgtcgcgact aagaaaatgc cgtcaaatcc gctcgccatg acttcactaa cgatgccttt
     4741 gaaaatcttc aagttctttt ctactaattc aaggcgtgtc tcaccaggtt tttggtttgc
     4801 tccggcgcaa atgcagacaa tatcagcatc cttgcagggt atgtttctct ttgatgtctt
     4861 tttgtttgtg aagtatttca catttatatt gtgcaacact tcacaaactt ttgcaagaga
     4921 aaagttttgt ctgatttatg aacaaaaaag aaaccatcat tgatggtttc tttcggtaag
     4981 tcccgtctag ccttgccctc aatggggaag agaaccgctt aagcccgagt cattatataa
     5041 accatttagc acgtaatcaa agccaggctg attctgaccg ggcacttggg cgctgccatt
     5101 attaaaaatc acttttgcgt tggttgtatc cgtgtccgca ggcagcgtca gcgtgtaaat
     5161 tccgtctgca tttttagtca ttggttttcc aggccaagat ccggtcaatt caattactcg
     5221 gctcccatca tgtttataga tataagcatt tacctggctc caatgattcg gattttgata
     5281 gccgatggtt ttggccgacg ctggatctct tttaacaaaa ctgtatttct cggtcctcgt
     5341 tacaccatca ctgttcgttc cttttaacat gatggtgtat gttttgccaa atggatctcc
     5401 ttttccgatt gtgaattgat ctccatcctt aaacgcctgt cgtctggtcc attattgatt
     5461 tgataaacgg cttttgttgt attcgcatct gcacgcaagg taatcgtcag ttgatcattg
     5521 aaagaatgtg ttacacctgt tttgtaattc tcaaggaaaa catgaggcgc ttttgcaata
     5581 tcatcaggat aaagcacagc tacagacctg gcattgatcg tgcctgtcag tttaccatcg
     5641 ttcacttgaa atgaacccgc tccagcttta ttgtcatacc tgccatcagg caattttgtt
     5701 gccgtattga tagagacaga ggatgaacct gcatttgcca gcacaacgcc atgtgagccg
     5761 cgctgattca taaatatctg gttgtttcca ttcgggttcg agagttcctc aggctgtcca
     5821 gccatcacat tgtgaaatct attgaccgca gtgatagcct gatcttcaaa taaagcactc
     5881 ccgcgatcgc ctatttggct tttccccggg aacctcacac catttccgcc tccctcaggt
     5941 ctggaaaaga aaagaggcgt actgcctgaa cgagaagcta tcaccgccca gcctaaacgg
     6001 attcccccta tgcaagggtt tattgttttc taaaatctga ttaccaatta gaatgaatat
     6061 ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa gtgtttccac cattttttca
     6121 atttttttat aattttttta atctgttatt taaatagttt atagttaaat ttacattttc
     6181 attagtccat tcaatattct ctccaagata actacgaact gctaacaaaa ttctctccct
     6241 atgttctaat ggagaagatt cagccactgc atttcccgca atatcttttg gtatgatttt
     6301 acccgtgtcc atagttaaaa tcatacggca taaagttaat atagagttgg tttcatcatc
     6361 ctgataatta tctattaatt cctctgacga atccataatg gctcttctca catcagaaaa
     6421 tggaatatca ggtagtaatt cctctaagtc ataatttccg tatattcttt tattttttcg
     6481 ttttgcttgg taaagcatta tggttaaatc tgaatttaat tccttctgag gaatgtatcc
     6541 ttgttcataa agctcttgta accattctcc ataaataaat tcttgtttgg gaggatgatt
     6601 ccacggtacc atttcttgct gaataataat tgttaattca atatatcgta agttgctttt
     6661 atctcctatt ttttttgaaa taggtctaat tttttgtata agtatttctt tactttgatc
     6721 tgtcaatggt tcagatacga cgactaaaaa gtcaagatca ctatttggtt ttagtccact
     6781 ctcaactcct gatccaaaca tgtaagtacc aataaggtta ttttttaaat gtttccgaag
     6841 tatttttttc actttattaa tttgttcgta tgtattcaaa tatatcctcc tcactatttt
     6901 gattagtacc tattttatat ccatagttgt taattaaata aacttaattt agtttattta
     6961 tagatttcat tggcttctaa attttttatc tagataataa ttattttagt taattttatt
     7021 ctagattata tatgatatga tctttcattt ccataaaact aaagtaagtg taaacctatt
     7081 cattgtttta aaaatatctc ttgccagtca cgttacgtta ttagttatag ttattataac
     7141 atgtattcac gaacgaaaat cgccattcgc cagctgcagg aattcgatat caagcttcta
     7201 ggatctttac attttaacga tatctagaaa atgttcaata acaatggaag acattcctaa
     7261 tgccggtgca ttctttccta aggaagatgg taataattca tagctattgc ctaattgggg
     7321 ataaaccctg gacgacactt cacttctaat tgaatttaaa accataggat gtgattcaat
     7381 tatgctgttt cttaaaatga tggcttgtgg attgaatgta tttagaatat tagtaaggcc
     7441 aattcctaaa tagaatccga aattctgtaa tgcatttaag gtgccgatat cattcagatg
     7501 ggcgaggtct atgatatctt gataggacac ttttttctct ttagtctgaa gagattttaa
     7561 taaagccttc tctgaagcat acaattccca gcaacctcgg tttccgcaac tgcatttagg
     7621 accattaaag tctattgtca tatgtcccat ttctccagag aatccgctta cccctctata
     7681 taaatgattg ttgataataa caccgatccc tatacctgta ctgatactag cataaataat
     7741 gttattgtga ttttttgcag caccaaatac tttttctcca tacgcgccag catttgcctc
     7801 attttcaata aaaacaggca cattgaactt ctcttgtatg aaagatttta agtcaatatc
     7861 tctccagttg gagttcggag tgaaaacaat tttttgattt ttatcaatta gtccaggcac
     7921 gcatatgcct ataccgataa gcccgtacgg agattgtggc atacgcgtaa taaagtgatg
     7981 aatcatatca attaaaatgt ctttagttat ttctggagaa ttcgattcta aatggtgatg
     8041 ttgatcaaga atgatcgttc cttcaaggtc tgttaaaatg ccactaatat aatccacacc
     8101 aacatctatt cctatggagt atcctgcctt tttattaaaa acaagcatga caggtcttct
     8161 tccgccactt tgctgatcaa gaatgatcgt cccttcaagg tctgttaaaa tcccactaat
     8221 ataatccaca ccaacatcta tgcctatgga gtatcctgcc ttcttattaa aaacaagcat
     8281 cacaggtctt cttccgccac ttgattgtcc ttgacctatt tcaaatacaa gattttcttt
     8341 catcagcgtg tttacctgtg atgaaacagt tgatttattt aatcc
//
