LOCUS       sequence-252406-        7376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gfpmut2 constitutively expressed using the Ppen promoter for 
            expression in B. subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7376)
  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 7376)
  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: pLG166
FEATURES             Location/Qualifiers
     source          1..7376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      66..654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     555..574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4165..4251
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4270..4297
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4377..4411
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             4573..5289
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKY"
     primer_bind     complement(4600..4628)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5223..5243
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6220..6238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6306..6410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6411..6479,6480..7271)
                     /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(6629..6648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
       61 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      181 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
      241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      661 agcaacgcgg cccgacctcg agctggatac ttcccgtccg ccagggggac atgccggcga
      721 tgctgaaggt cgcgcgcatt cccgatgaag aggccggtta ccgcctgttt gaggatatag
      781 taatctttct aaatagcttt ggattggagg agtatggcca ctaatactaa gttcagctaa
      841 taaaaaaatt tgctaaagaa ctccagctgg atttcactga tgagaatatc gtcggagata
      901 aatataataa ttccacggac tatagactat actagtatac tccgtctact gtacgataca
      961 cttccgctca ggtccttgtc ctttaacgag gattgttacg tacgctaatg gcgtcaaaac
     1021 aaagactcta gacctaggcc ttaagatctg atcatatgca tccgcgggcc cgggttaacg
     1081 cgtaatccat ggatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     1141 tgcacgcagg ttctccggtg ccctgaatga actgcagaaa gagctggtag ttggcgcact
     1201 gttcgaagaa ctgccgatgt ccagtaagat tcttactatg ctggttgaac cggatgctgg
     1261 taaagctact tgggttgctg cttctactta tggtaccgat acaactactg gtgaggaagt
     1321 taaaggagct cttaaagaaa tccacttcag taggtggccc ggctccatgc accgcgacgc
     1381 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     1441 gtgctcgccg aggcggtata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     1501 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     1561 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     1621 gggaaggcca tccagcctcg cgtcgcgact aagaaaatgc cgtcaaatcc gctcgccatg
     1681 acttcactaa cgatgccttt gaaaatcttc aagttctttt ctactaattc aaggcgtgtc
     1741 tcaccaggtt tttggtttgc tccggcgcaa atgcagacaa tatcagcatc cttgcagggt
     1801 atgtttctct ttgatgtctt tttgtttgtg aagtatttca catttatatt gtgcaacact
     1861 tcacaaactt ttgcaagaga aaagttttgt ctgatttatg aacaaaaaag aaaccatcat
     1921 tgatggtttc tttcggtaag tcccgtctag ccttgccctc aatggggaag agaaccgctt
     1981 aagcccgagt cattatataa accatttagc acgtaatcaa agccaggctg attctgaccg
     2041 ggcacttggg cgctgccatt attaaaaatc acttttgcgt tggttgtatc cgtgtccgca
     2101 ggcagcgtca gcgtgtaaat tccgtctgca tttttagtca ttggttttcc aggccaagat
     2161 ccggtcaatt caattactcg gctcccatca tgtttataga tataagcatt tacctggctc
     2221 caatgattcg gattttgata gccgatggtt ttggccgacg ctggatctct tttaacaaaa
     2281 ctgtatttct cggtcctcgt tacaccatca ctgttcgttc cttttaacat gatggtgtat
     2341 gttttgccaa atggatctcc ttttccgatt gtgaattgat ctccatcctt aaacgcctgt
     2401 cgtctggtcc attattgatt tgataaacgg cttttgttgt attcgcatct gcacgcaagg
     2461 taatcgtcag ttgatcattg aaagaatgtg ttacacctgt tttgtaattc tcaaggaaaa
     2521 catgaggcgc ttttgcaata tcatcaggat aaagcacagc tacagacctg gcattgatcg
     2581 tgcctgtcag tttaccatcg ttcacttgaa atgaacccgc tccagcttta ttgtcatacc
     2641 tgccatcagg caattttgtt gccgtattga tagagacaga ggatgaacct gcatttgcca
     2701 gcacaacgcc atgtgagccg cgctgattca taaatatctg gttgtttcca ttcgggttcg
     2761 agagttcctc aggctgtcca gccatcacat tgtgaaatct attgaccgca gtgatagcct
     2821 gatcttcaaa taaagcactc ccgcgatcgc ctatttggct tttccccggg aacctcacac
     2881 catttccgcc tccctcaggt ctggaaaaga aaagaggcgt actgcctgaa cgagaagcta
     2941 tcaccgccca gcctaaacgg attcccccta tgcaagggtt tattgttttc taaaatctga
     3001 ttaccaatta gaatgaatat ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa
     3061 gtgtttccac cattttttca atttttttat aattttttta atctgttatt taaatagttt
     3121 atagttaaat ttacattttc attagtccat tcaatattct ctccaagata actacgaact
     3181 gctaacaaaa ttctctccct atgttctaat ggagaagatt cagccactgc atttcccgca
     3241 atatcttttg gtatgatttt acccgtgtcc atagttaaaa tcatacggca taaagttaat
     3301 atagagttgg tttcatcatc ctgataatta tctattaatt cctctgacga atccataatg
     3361 gctcttctca catcagaaaa tggaatatca ggtagtaatt cctctaagtc ataatttccg
     3421 tatattcttt tattttttcg ttttgcttgg taaagcatta tggttaaatc tgaatttaat
     3481 tccttctgag gaatgtatcc ttgttcataa agctcttgta accattctcc ataaataaat
     3541 tcttgtttgg gaggatgatt ccacggtacc atttcttgct gaataataat tgttaattca
     3601 atatatcgta agttgctttt atctcctatt ttttttgaaa taggtctaat tttttgtata
     3661 agtatttctt tactttgatc tgtcaatggt tcagatacga cgactaaaaa gtcaagatca
     3721 ctatttggtt ttagtccact ctcaactcct gatccaaaca tgtaagtacc aataaggtta
     3781 ttttttaaat gtttccgaag tatttttttc actttattaa tttgttcgta tgtattcaaa
     3841 tatatcctcc tcactatttt gattagtacc tattttatat ccatagttgt taattaaata
     3901 aacttaattt agtttattta tagatttcat tggcttctaa attttttatc tagataataa
     3961 ttattttagt taattttatt ctagattata tatgatatga tctttcattt ccataaaact
     4021 aaagtaagtg taaacctatt cattgtttta aaaatatctc ttgccagtca cgttacgtta
     4081 ttagttatag ttattataac atgtattcac gaacgaaaat cgccattcgc cagctgcagg
     4141 aattcgacct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4201 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgctc
     4261 tagctaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttgtta
     4321 actctagagc tgcctgccgc gtttcggtga tgaagatctt cccgatgatt aattaattca
     4381 gaacgctcgg ttgccgccgg gcgtttttta tgcagcaatg gcaagaacgt tgctcgaggg
     4441 taactcggtg gaaacgaggt catcatttcc ttccgaaaaa acggttgcat ttaaatctta
     4501 catatgtaat actttcaaag actacatttg taagatttga tgtttgagtc ggctgaaagg
     4561 tggtgaacta ctatgaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     4621 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     4681 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     4741 ccaacacttg tcactacttt cgcgtatggt cttcaatgct ttgcgagata cccagatcat
     4801 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
     4861 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     4921 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     4981 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     5041 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     5101 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     5161 aaccattacc tgtccacaca atctaagctt tcgaaagatc ccaacgaaaa gagggaccac
     5221 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     5281 aaatactagc tgcattcagc ccgcctaatg agcgggcttt tttaattcaa tttcacaatc
     5341 agagagaacg aattcgataa ccctaaagtt atggaaataa gacttagaag caaacttaag
     5401 agtgtgttga tagtgcatta tcttaaaatt ttgtataata ggaattgaag ttaaattaga
     5461 tgctaaaaat ttgtaattaa gaaggaggga ttcgtcatgt tggggagccc gtatttcgcg
     5521 taaggaaatc cattatgtac tatttaattc tgcgtgacat cccatcgatc agaccagttt
     5581 ttaatttgtg tgtttccatg tgtccagttt ggaatactct taacctcatt ggaaatcgcg
     5641 gcataatcac tggtggtatg attgatgacc gcgtcaacaa tgacctttat gccatattct
     5701 tcagcggctg cacacatttc tttaaattct tgttcagtac ctaagtaacg gttgccaatt
     5761 tgatacgatg tcggctgata cagccagtac cagttcgaca tgcttttatc tccttgattc
     5821 ccttccttta cttggttaat cggagatgtc tgaatggctg tatatcctgc atcatgaata
     5881 tccttcatat tgtgttttaa cgtattgaac gaccaattcc atgcatgaag aatggttccg
     5941 cttttgatcg acggtgctgt aagctcattc gatttgttcg ccgtttcagc actcgcagcc
     6001 gccggtcctg ccagaaccaa atgaaacagc aataaaaatc cagcgaataa cggcagtaaa
     6061 gaggttttga atcgttttgc aaacattctt gacactcctt atttgatttt ttgaagactt
     6121 acttcggagt caaaaatccc tcttacttca ttcttccgct tcctcctttc aaaccgatgt
     6181 gaagactgga gaattttgtt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     6241 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6301 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6361 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6421 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6481 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6541 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6601 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     6661 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6721 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6781 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6841 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6901 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     6961 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7021 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7081 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtcg cgcggtatca
     7141 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7201 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7261 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7321 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaa
//
