LOCUS       sequence-252487-        8491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vanillic acid inducible production of lysostaphin. LsyT with the 
            amyE signaling peptide for secretion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8491)
  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 8491)
  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: pNM262
FEATURES             Location/Qualifiers
     source          1..8491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(769..787)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        855..959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(960..1028,1029..1820)
                     /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(1178..1197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1991..2579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2480..2499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6085..6171
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6190..6217
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        6404..6453
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      7376..7470
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg
       61 tatttcgcgt aaggaaatcc attatgtact atttaattct gcgtgacatc ccatcgatca
      121 gaccagtttt taatttgtgt gtttccatgt gtccagtttg gaatactctt aacctcattg
      181 gaaatcgcgg cataatcact ggtggtatga ttgatgaccg cgtcaacaat gacctttatg
      241 ccatattctt cagcggctgc acacatttct ttaaattctt gttcagtacc taagtaacgg
      301 ttgccaattt gatacgatgt cggctgatac agccagtacc agttcgacat gcttttatct
      361 ccttgattcc cttcctttac ttggttaatc ggagatgtct gaatggctgt atatcctgca
      421 tcatgaatat ccttcatatt gtgttttaac gtattgaacg accaattcca tgcatgaaga
      481 atggttccgc ttttgatcga cggtgctgta agctcattcg atttgttcgc cgtttcagca
      541 ctcgcagccg ccggtcctgc cagaaccaaa tgaaacagca ataaaaatcc agcgaataac
      601 ggcagtaaag aggttttgaa tcgttttgca aacattcttg acactcctta tttgattttt
      661 tgaagactta cttcggagtc aaaaatccct cttacttcat tcttccgctt cctcctttca
      721 aaccgatgtg aagactggag aattttgtta attcttgaag acgaaagggc ctcgtgatac
      781 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
      841 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      901 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      961 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1021 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1081 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1141 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1201 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1261 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1321 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1381 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1441 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1501 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1561 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1621 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtcgc
     1681 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1741 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1801 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1861 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2101 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2581 acgccagcaa cgcggcccga cctcgagctg gatacttccc gtccgccagg gggacatgcc
     2641 ggcgatgctg aaggtcgcgc gcattcccga tgaagaggcc ggttaccgcc tgtttgagga
     2701 tatagtaatc tttctaaata gctttggatt ggaggagtat ggccactaat actaagttca
     2761 gctaataaaa aaatttgcta aagaactcca gctggatttc actgatgaga atatcgtcgg
     2821 agataaatat aataattcca ctatagacta tactagtata ctccgtctac tgtacgatac
     2881 acttccgctc aggtccttgt cctttaacga ggattgttac gtacgctaat ggcgtcaaaa
     2941 caaagactct agacctaggc cttaagatct gatcatatgc atccgcgggc ccgggttaac
     3001 gcgtaatcca tggatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3061 ttgcacgcag gttctccggt gccctgaatg aactgcagaa agagctggta gttggcgcac
     3121 tgttcgaaga actgccgatg tccagtaaga ttcttactat gctggttgaa ccggatgctg
     3181 gtaaagctac ttgggttgct gcttctactt atggtaccga tacaactact ggtgaggaag
     3241 ttaaaggagc tcttaaagaa atccacttca gtaggtggcc cggctccatg caccgcgacg
     3301 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
     3361 tgtgctcgcc gaggcggtat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
     3421 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
     3481 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
     3541 ggggaaggcc atccagcctc gcgtcgcgac taagaaaatg ccgtcaaatc cgctcgccat
     3601 gacttcacta acgatgcctt tgaaaatctt caagttcttt tctactaatt caaggcgtgt
     3661 ctcaccaggt ttttggtttg ctccggcgca aatgcagaca atatcagcat ccttgcaggg
     3721 tatgtttctc tttgatgtct ttttgtttgt gaagtatttc acatttatat tgtgcaacac
     3781 ttcacaaact tttgcaagag aaaagttttg tctgatttat gaacaaaaaa gaaaccatca
     3841 ttgatggttt ctttcggtaa gtcccgtcta gccttgccct caatggggaa gagaaccgct
     3901 taagcccgag tcattatata aaccatttag cacgtaatca aagccaggct gattctgacc
     3961 gggcacttgg gcgctgccat tattaaaaat cacttttgcg ttggttgtat ccgtgtccgc
     4021 aggcagcgtc agcgtgtaaa ttccgtctgc atttttagtc attggttttc caggccaaga
     4081 tccggtcaat tcaattactc ggctcccatc atgtttatag atataagcat ttcctggctc
     4141 caatgattcg gattttgata gccgatggtt ttggccgacg ctggatctct tttaacaaaa
     4201 ctgtatttct cggtcctcgt tacaccatca ctgttcgttc cttttaacat gatggtgtat
     4261 gttttgccaa atggatctcc ttttccgatt gtgaattgat ctccatcctt aaacgcctgt
     4321 cgtctggtcc attattgatt tgataaacgg cttttgttgt attcgcatct gcacgcaagg
     4381 taatcgtcag ttgatcattg aaagaatgtg ttacacctgt tttgtaattc tcaaggaaaa
     4441 catgaggcgc ttttgcaata tcatcaggat aaagcacagc tacagacctg gcattgatcg
     4501 tgcctgtcag tttaccatcg ttcacttgaa atgaacccgc tccagcttta ttgtcatacc
     4561 tgccatcagg caattttgtt gccgtattga tagagacaga ggatgaacct gcatttgcca
     4621 gcacaacgcc atgtgagccg cgctgattca taaatatctg gttgtttcca ttcgggttcg
     4681 agagttcctc aggctgtcca gccatcacat tgtgaaatct attgaccgca gtgatagcct
     4741 gatcttcaaa taaagcactc ccgcgatcgc ctatttggct tttccccggg aacctcacac
     4801 catttccgcc tccctcaggt ctggaaaaga aaagaggcgt actgcctgaa cgagaagcta
     4861 tcaccgccca gcctaaacgg attcccccta tgcaagggtt tattgttttc taaaatctga
     4921 ttaccaatta gaatgaatat ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa
     4981 gtgtttccac cattttttca atttttttat aattttttta atctgttatt taaatagttt
     5041 atagttaaat ttacattttc attagtccat tcaatattct ctccaagata actacgaact
     5101 gctaacaaaa ttctctccct atgttctaat ggagaagatt cagccactgc atttcccgca
     5161 atatcttttg gtatgatttt acccgtgtcc atagttaaaa tcatacggca taaagttaat
     5221 atagagttgg tttcatcatc ctgataatta tctattaatt cctctgacga atccataatg
     5281 gctcttctca catcagaaaa tggaatatca ggtagtaatt cctctaagtc ataatttccg
     5341 tatattcttt tattttttcg ttttgcttgg taaagcatta tggttaaatc tgaatttaat
     5401 tccttctgag gaatgtatcc ttgttcataa agctcttgta accattctcc ataaataaat
     5461 tcttgtttgg gaggatgatt ccacggtacc atttcttgct gaataataat tgttaattca
     5521 atatatcgta agttgctttt atctcctatt ttttttgaaa taggtctaat tttttgtata
     5581 agtatttctt tactttgatc tgtcaatggt tcagatacga cgactaaaaa gtcaagatca
     5641 ctatttggtt ttagtccact ctcaactcct gatccaaaca tgtaagtacc aataaggtta
     5701 ttttttaaat gtttccgaag tatttttttc actttattaa tttgttcgta tgtattcaaa
     5761 tatatcctcc tcactatttt gattagtacc tattttatat ccatagttgt taattaaata
     5821 aacttaattt agtttattta tagatttcat tggcttctaa attttttatc tagataataa
     5881 ttattttagt taattttatt ctagattata tatgatatga tctttcattt ccataaaact
     5941 aaagtaagtg taaacctatt cattgtttta aaaatatctc ttgccagtca cgttacgtta
     6001 ttagttatag ttattataac atgtattcac gaacgaaaat cgccattcgc cagctgcagg
     6061 aattcgacct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     6121 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgctc
     6181 tagctaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttgtta
     6241 actctagagc tgcctgccgc gtttcggtga tgaagatctt cccgatgatt attaattcag
     6301 aacgctcggt tgccgcggca agaacgttgc tcgagattgg atccaatcat tttgcaaaag
     6361 ttgttgactt tatctacaag gtgtggcata ttggatccaa tagctgtcac cggatgtgct
     6421 ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt gtttaatcgg
     6481 ctgtaaagga ggaattcaaa atgtttgcaa aacgattcaa aacctcttta ctgccgttat
     6541 tcgctggatt tttattgctg tttcatttgg ttctggcagg accggcggct gcgagtgctg
     6601 gaagcacaca tgaacattca gcacaatggt tgaataatta caaaaaagga tatggttacg
     6661 gtccttatcc attaggtata aatggcggta tgcactacgg agttgatttt tttatgaata
     6721 ttggaacacc agtaaaagct atttcaagcg gaaaaatagt tgaagctggt tggagtaatt
     6781 acggaggagg taatcaaata ggtcttattg aaaatgatgg agtgcataga caatggtata
     6841 tgcatctaag taaatataat gttaaagtag gagattatgt caaagctggt caaataatcg
     6901 gttggtctgg aagcactggt tattctacag caccacattt acacttccaa agaatggtta
     6961 attcattttc aaattcaact gcccaagatc caatgccttt cttaaagagc gcaggatatg
     7021 gaaaagcagg tggtacagta actccaacgc cgaatacagg ttggaaaaca aacaaatatg
     7081 gcacactata taaatcagag tcagctagct tcacacctaa tacagatata ataacaagaa
     7141 cgactggtcc atttagaagc atgccgcagt caggagtctt aaaagcaggt caaacaattc
     7201 attatgatga agtgatgaaa caagacggtc atgtttgggt aggttataca ggtaacagtg
     7261 gccaacgtat ttacttgcct gtaagaacat ggaataaatc tactaatact ttaggtgttc
     7321 tttggggaac tataaagtaa cgtcgtgact gggaaaaccc tggcgactag tcttggactc
     7381 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     7441 tgccgccggg cgttttttat tggtgagaat ccaggggtcc ccaataatta cgatttaaat
     7501 tggagcttag tcgacagcta gccgcatgca agctaattcg gtggaaacga ggtcatcatt
     7561 tccttccgaa aaaacggttg catttaaatc ttacatatgt aatactttca aagactacat
     7621 ttgtaagatt tgatgtttgt agcgcccgtc ggctgaaagg tggtgaacta ctatggacat
     7681 gcctcgtatt aaaccgggtc agcgtgttat gatggcactg cgtaaaatga ttgcaagcgg
     7741 tgaaatcaaa agtggtgaac gtattgcaga aattccgacc gcagcagcac tgggtgttag
     7801 ccgtatgccg gttcgtatcg cactgcgttc actggaacaa gaaggtctgg ttgttcgtct
     7861 gggtgcacgt ggttatgcag cccgtggtgt tagcagcgat cagattcgtg atgcaattga
     7921 agttcgtggt gttctggaag gttttgcagc acgtcgtctg gcagaacgtg gtatgaccgc
     7981 agaaacccat gcacgttttg ttgtactgat tgcagaaggt gaagcactgt ttgcagccgg
     8041 tcgcctgaat ggtgaagatc tggatcgtta tgccgcatat aatcaggcat ttcatgatac
     8101 cctggttagc gcagcaggta atggtgcagt tgaaagcgca ctggcacgta atggttttga
     8161 accgtttgca gcagccggtg cactggccct ggatctgatg gacctgtctg ccgaatatga
     8221 acatctgctg gcagcacatc gtcagcatca ggcagttctg gatgcagtta gctgtggtga
     8281 tgccgaaggt gcagaacgta ttatgcgtga tcatgcactg gcagcaattc gtaatgcaaa
     8341 agtttttgaa gcagcagcaa gcgcaggcgc accgctgggt gcagcatggt caattcgtgc
     8401 agattgataa ggatcctaat tggtaacgaa tcagacaatt gacggcgcgc aacgcaatta
     8461 atgtgagtta ggatcctgag cgccggtcgc t
//
