LOCUS       sequence-256757-        8389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vanillic acid inducible production of lysostaphin. LsyT with no 
            signaling peptide for secretion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8389)
  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 8389)
  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: pNM261
FEATURES             Location/Qualifiers
     source          1..8389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      270..356
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      375..402
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        589..638
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      1459..1553
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3343..3361)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3429..3533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3534..3602,3603..4394)
                     /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(3752..3771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4565..5153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5054..5073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaataaactt aatttagttt atttatagat ttcattggct tctaaatttt ttatctagat
       61 aataattatt ttagttaatt ttattctaga ttatatatga tatgatcttt catttccata
      121 aaactaaagt aagtgtaaac ctattcattg ttttaaaaat atctcttgcc agtcacgtta
      181 cgttattagt tatagttatt ataacatgta ttcacgaacg aaaatcgcca ttcgccagct
      241 gcaggaattc gacctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact
      301 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      361 cgctctagct aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
      421 tgttaactct agagctgcct gccgcgtttc ggtgatgaag atcttcccga tgattattaa
      481 ttcagaacgc tcggttgccg cggcaagaac gttgctcgag attggatcca atcattttgc
      541 aaaagttgtt gactttatct acaaggtgtg gcatattgga tccaatagct gtcaccggat
      601 gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata attttgttta
      661 atcggctgta aaggaggaat tcaaaatgac acatgaacat tcagcacaat ggttgaataa
      721 ttacaaaaaa ggatatggtt acggtcctta tccattaggt ataaatggcg gtatgcacta
      781 cggagttgat ttttttatga atattggaac accagtaaaa gctatttcaa gcggaaaaat
      841 agttgaagct ggttggagta attacggagg aggtaatcaa ataggtctta ttgaaaatga
      901 tggagtgcat agacaatggt atatgcatct aagtaaatat aatgttaaag taggagatta
      961 tgtcaaagct ggtcaaataa tcggttggtc tggaagcact ggttattcta cagcaccaca
     1021 tttacacttc caaagaatgg ttaattcatt ttcaaattca actgcccaag atccaatgcc
     1081 tttcttaaag agcgcaggat atggaaaagc aggtggtaca gtaactccaa cgccgaatac
     1141 aggttggaaa acaaacaaat atggcacact atataaatca gagtcagcta gcttcacacc
     1201 taatacagat ataataacaa gaacgactgg tccatttaga agcatgccgc agtcaggagt
     1261 cttaaaagca ggtcaaacaa ttcattatga tgaagtgatg aaacaagacg gtcatgtttg
     1321 ggtaggttat acaggtaaca gtggccaacg tatttacttg cctgtaagaa catggaataa
     1381 atctactaat actttaggtg ttctttgggg aactataaag taacgtcgtg actgggaaaa
     1441 ccctggcgac tagtcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1501 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccagggg
     1561 tccccaataa ttacgattta aattggagct tagtcgacag ctagccgcat gcaagctaat
     1621 tcggtggaaa cgaggtcatc atttccttcc gaaaaaacgg ttgcatttaa atcttacata
     1681 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgtagcgccc gtcggctgaa
     1741 aggtggtgaa ctactatgga catgcctcgt attaaaccgg gtcagcgtgt tatgatggca
     1801 ctgcgtaaaa tgattgcaag cggtgaaatc aaaagtggtg aacgtattgc agaaattccg
     1861 accgcagcag cactgggtgt tagccgtatg ccggttcgta tcgcactgcg ttcactggaa
     1921 caagaaggtc tggttgttcg tctgggtgca cgtggttatg cagcccgtgg tgttagcagc
     1981 gatcagattc gtgatgcaat tgaagttcgt ggtgttctgg aaggttttgc agcacgtcgt
     2041 ctggcagaac gtggtatgac cgcagaaacc catgcacgtt ttgttgtact gattgcagaa
     2101 ggtgaagcac tgtttgcagc cggtcgcctg aatggtgaag atctggatcg ttatgccgca
     2161 tataatcagg catttcatga taccctggtt agcgcagcag gtaatggtgc agttgaaagc
     2221 gcactggcac gtaatggttt tgaaccgttt gcagcagccg gtgcactggc cctggatctg
     2281 atggacctgt ctgccgaata tgaacatctg ctggcagcac atcgtcagca tcaggcagtt
     2341 ctggatgcag ttagctgtgg tgatgccgaa ggtgcagaac gtattatgcg tgatcatgca
     2401 ctggcagcaa ttcgtaatgc aaaagttttt gaagcagcag caagcgcagg cgcaccgctg
     2461 ggtgcagcat ggtcaattcg tgcagattga taaggatcct aattggtaac gaatcagaca
     2521 attgacggcg cgcaacgcaa ttaatgtgag ttaggatcct gagcgccggt cgctaccatt
     2581 accagttggt ctggtgtcaa aaataataat aaccgggcag gccatgtctg cccgtatttc
     2641 gcgtaaggaa atccattatg tactatttaa ttctgcgtga catcccatcg atcagaccag
     2701 tttttaattt gtgtgtttcc atgtgtccag tttggaatac tcttaacctc attggaaatc
     2761 gcggcataat cactggtggt atgattgatg accgcgtcaa caatgacctt tatgccatat
     2821 tcttcagcgg ctgcacacat ttctttaaat tcttgttcag tacctaagta acggttgcca
     2881 atttgatacg atgtcggctg atacagccag taccagttcg acatgctttt atctccttga
     2941 ttcccttcct ttacttggtt aatcggagat gtctgaatgg ctgtatatcc tgcatcatga
     3001 atatccttca tattgtgttt taacgtattg aacgaccaat tccatgcatg aagaatggtt
     3061 ccgcttttga tcgacggtgc tgtaagctca ttcgatttgt tcgccgtttc agcactcgca
     3121 gccgccggtc ctgccagaac caaatgaaac agcaataaaa atccagcgaa taacggcagt
     3181 aaagaggttt tgaatcgttt tgcaaacatt cttgacactc cttatttgat tttttgaaga
     3241 cttacttcgg agtcaaaaat ccctcttact tcattcttcc gcttcctcct ttcaaaccga
     3301 tgtgaagact ggagaatttt gttaattctt gaagacgaaa gggcctcgtg atacgcctat
     3361 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3421 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3481 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3541 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3601 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3661 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3721 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     3781 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3841 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3901 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3961 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4021 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     4081 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4141 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4201 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcgcgcggta
     4261 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4321 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4381 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4441 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4501 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4561 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4621 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4681 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4741 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4801 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4861 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4921 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4981 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5041 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5101 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5161 gcaacgcggc ccgacctcga gctggatact tcccgtccgc cagggggaca tgccggcgat
     5221 gctgaaggtc gcgcgcattc ccgatgaaga ggccggttac cgcctgtttg aggatatagt
     5281 aatctttcta aatagctttg gattggagga gtatggccac taatactaag ttcagctaat
     5341 aaaaaaattt gctaaagaac tccagctgga tttcactgat gagaatatcg tcggagataa
     5401 atataataat tccactatag actatactag tatactccgt ctactgtacg atacacttcc
     5461 gctcaggtcc ttgtccttta acgaggattg ttacgtacgc taatggcgtc aaaacaaaga
     5521 ctctagacct aggccttaag atctgatcat atgcatccgc gggcccgggt taacgcgtaa
     5581 tccatggatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5641 gcaggttctc cggtgccctg aatgaactgc agaaagagct ggtagttggc gcactgttcg
     5701 aagaactgcc gatgtccagt aagattctta ctatgctggt tgaaccggat gctggtaaag
     5761 ctacttgggt tgctgcttct acttatggta ccgatacaac tactggtgag gaagttaaag
     5821 gagctcttaa agaaatccac ttcagtaggt ggcccggctc catgcaccgc gacgcaacgc
     5881 ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct
     5941 cgccgaggcg gtataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt
     6001 aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag
     6061 catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa
     6121 ggccatccag cctcgcgtcg cgactaagaa aatgccgtca aatccgctcg ccatgacttc
     6181 actaacgatg cctttgaaaa tcttcaagtt cttttctact aattcaaggc gtgtctcacc
     6241 aggtttttgg tttgctccgg cgcaaatgca gacaatatca gcatccttgc agggtatgtt
     6301 tctctttgat gtctttttgt ttgtgaagta tttcacattt atattgtgca acacttcaca
     6361 aacttttgca agagaaaagt tttgtctgat ttatgaacaa aaaagaaacc atcattgatg
     6421 gtttctttcg gtaagtcccg tctagccttg ccctcaatgg ggaagagaac cgcttaagcc
     6481 cgagtcatta tataaaccat ttagcacgta atcaaagcca ggctgattct gaccgggcac
     6541 ttgggcgctg ccattattaa aaatcacttt tgcgttggtt gtatccgtgt ccgcaggcag
     6601 cgtcagcgtg taaattccgt ctgcattttt agtcattggt tttccaggcc aagatccggt
     6661 caattcaatt actcggctcc catcatgttt atagatataa gcatttcctg gctccaatga
     6721 ttcggatttt gatagccgat ggttttggcc gacgctggat ctcttttaac aaaactgtat
     6781 ttctcggtcc tcgttacacc atcactgttc gttcctttta acatgatggt gtatgttttg
     6841 ccaaatggat ctccttttcc gattgtgaat tgatctccat ccttaaacgc ctgtcgtctg
     6901 gtccattatt gatttgataa acggcttttg ttgtattcgc atctgcacgc aaggtaatcg
     6961 tcagttgatc attgaaagaa tgtgttacac ctgttttgta attctcaagg aaaacatgag
     7021 gcgcttttgc aatatcatca ggataaagca cagctacaga cctggcattg atcgtgcctg
     7081 tcagtttacc atcgttcact tgaaatgaac ccgctccagc tttattgtca tacctgccat
     7141 caggcaattt tgttgccgta ttgatagaga cagaggatga acctgcattt gccagcacaa
     7201 cgccatgtga gccgcgctga ttcataaata tctggttgtt tccattcggg ttcgagagtt
     7261 cctcaggctg tccagccatc acattgtgaa atctattgac cgcagtgata gcctgatctt
     7321 caaataaagc actcccgcga tcgcctattt ggcttttccc cgggaacctc acaccatttc
     7381 cgcctccctc aggtctggaa aagaaaagag gcgtactgcc tgaacgagaa gctatcaccg
     7441 cccagcctaa acggattccc cctatgcaag ggtttattgt tttctaaaat ctgattacca
     7501 attagaatga atatttccca aatattaaat aataaaacaa aaaaattgaa aaaagtgttt
     7561 ccaccatttt ttcaattttt ttataatttt tttaatctgt tatttaaata gtttatagtt
     7621 aaatttacat tttcattagt ccattcaata ttctctccaa gataactacg aactgctaac
     7681 aaaattctct ccctatgttc taatggagaa gattcagcca ctgcatttcc cgcaatatct
     7741 tttggtatga ttttacccgt gtccatagtt aaaatcatac ggcataaagt taatatagag
     7801 ttggtttcat catcctgata attatctatt aattcctctg acgaatccat aatggctctt
     7861 ctcacatcag aaaatggaat atcaggtagt aattcctcta agtcataatt tccgtatatt
     7921 cttttatttt ttcgttttgc ttggtaaagc attatggtta aatctgaatt taattccttc
     7981 tgaggaatgt atccttgttc ataaagctct tgtaaccatt ctccataaat aaattcttgt
     8041 ttgggaggat gattccacgg taccatttct tgctgaataa taattgttaa ttcaatatat
     8101 cgtaagttgc ttttatctcc tatttttttt gaaataggtc taattttttg tataagtatt
     8161 tctttacttt gatctgtcaa tggttcagat acgacgacta aaaagtcaag atcactattt
     8221 ggttttagtc cactctcaac tcctgatcca aacatgtaag taccaataag gttatttttt
     8281 aaatgtttcc gaagtatttt tttcacttta ttaatttgtt cgtatgtatt caaatatatc
     8341 ctcctcacta ttttgattag tacctatttt atatccatag ttgttaatt
//
