LOCUS       sequence-246951-        8395 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 8395)
  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 8395)
  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..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..38)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        106..210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(211..279,280..1071)
                     /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(429..448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1242..1830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5341..5427
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5446..5473
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        5661..5710
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      6531..6625
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
       61 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      421 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
      481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      721 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
      781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
      901 gataaatctg gagccggtga gcgtgggtcg cgcggtatca ttgcagcact ggggccagat
      961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcccg acctcgagct
     1861 ggatacttcc cgtccgccag ggggacatgc cggcgatgct gaaggtcgcg cgcattcccg
     1921 atgaagaggc cggttaccgc ctgtttgagg atatagtaat ctttctaaat agctttggat
     1981 tggaggagta tggccactaa tactaagttc agctaataaa aaaatttgct aaagaactcc
     2041 agctggattt cactgatgag aatatcgtcg gagataaata taataattcc acggactata
     2101 gactatacta gtatactccg tctactgtac gatacacttc cgctcaggtc cttgtccttt
     2161 aacgaggatt gttacgtacg ctaatggcgt caaaacaaag actctagacc taggccttaa
     2221 gatctgatca tatgcatccg cgggcccggg ttaacgcgta atccatggat caagagacag
     2281 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggtgccct
     2341 gaatgaactg cagaaagagc tggtagttgg cgcactgttc gaagaactgc cgatgtccag
     2401 taagattctt actatgctgg ttgaaccgga tgctggtaaa gctacttggg ttgctgcttc
     2461 tacttatggt accgatacaa ctactggtga ggaagttaaa ggagctctta aagaaatcca
     2521 cttcagtagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     2581 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     2641 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     2701 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     2761 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     2821 gcgactaaga aaatgccgtc aaatccgctc gccatgactt cactaacgat gcctttgaaa
     2881 atcttcaagt tcttttctac taattcaagg cgtgtctcac caggtttttg gtttgctccg
     2941 gcgcaaatgc agacaatatc agcatccttg cagggtatgt ttctctttga tgtctttttg
     3001 tttgtgaagt atttcacatt tatattgtgc aacacttcac aaacttttgc aagagaaaag
     3061 ttttgtctga tttatgaaca aaaaagaaac catcattgat ggtttctttc ggtaagtccc
     3121 gtctagcctt gccctcaatg gggaagagaa ccgcttaagc ccgagtcatt atataaacca
     3181 tttagcacgt aatcaaagcc aggctgattc tgaccgggca cttgggcgct gccattatta
     3241 aaaatcactt ttgcgttggt tgtatccgtg tccgcaggca gcgtcagcgt gtaaattccg
     3301 tctgcatttt tagtcattgg ttttccaggc caagatccgg tcaattcaat tactcggctc
     3361 ccatcatgtt tatagatata agcatttacc tggctccaat gattcggatt ttgatagccg
     3421 atggttttgg ccgacgctgg atctctttta acaaaactgt atttctcggt cctcgttaca
     3481 ccatcactgt tcgttccttt taacatgatg gtgtatgttt tgccaaatgg atctcctttt
     3541 ccgattgtga attgatctcc atccttaaac gcctgtcgtc tggtccatta ttgatttgat
     3601 aaacggcttt tgttgtattc gcatctgcac gcaaggtaat cgtcagttga tcattgaaag
     3661 aatgtgttac acctgttttg taattctcaa ggaaaacatg aggcgctttt gcaatatcat
     3721 caggataaag cacagctaca gacctggcat tgatcgtgcc tgtcagttta ccatcgttca
     3781 cttgaaatga acccgctcca gctttattgt catacctgcc atcaggcaat tttgttgccg
     3841 tattgataga gacagaggat gaacctgcat ttgccagcac aacgccatgt gagccgcgct
     3901 gattcataaa tatctggttg tttccattcg ggttcgagag ttcctcaggc tgtccagcca
     3961 tcacattgtg aaatctattg accgcagtga tagcctgatc ttcaaataaa gcactcccgc
     4021 gatcgcctat ttggcttttc cccgggaacc tcacaccatt tccgcctccc tcaggtctgg
     4081 aaaagaaaag aggcgtactg cctgaacgag aagctatcac cgcccagcct aaacggattc
     4141 cccctatgca agggtttatt gttttctaaa atctgattac caattagaat gaatatttcc
     4201 caaatattaa ataataaaac aaaaaaattg aaaaaagtgt ttccaccatt ttttcaattt
     4261 ttttataatt tttttaatct gttatttaaa tagtttatag ttaaatttac attttcatta
     4321 gtccattcaa tattctctcc aagataacta cgaactgcta acaaaattct ctccctatgt
     4381 tctaatggag aagattcagc cactgcattt cccgcaatat cttttggtat gattttaccc
     4441 gtgtccatag ttaaaatcat acggcataaa gttaatatag agttggtttc atcatcctga
     4501 taattatcta ttaattcctc tgacgaatcc ataatggctc ttctcacatc agaaaatgga
     4561 atatcaggta gtaattcctc taagtcataa tttccgtata ttcttttatt ttttcgtttt
     4621 gcttggtaaa gcattatggt taaatctgaa tttaattcct tctgaggaat gtatccttgt
     4681 tcataaagct cttgtaacca ttctccataa ataaattctt gtttgggagg atgattccac
     4741 ggtaccattt cttgctgaat aataattgtt aattcaatat atcgtaagtt gcttttatct
     4801 cctatttttt ttgaaatagg tctaattttt tgtataagta tttctttact ttgatctgtc
     4861 aatggttcag atacgacgac taaaaagtca agatcactat ttggttttag tccactctca
     4921 actcctgatc caaacatgta agtaccaata aggttatttt ttaaatgttt ccgaagtatt
     4981 tttttcactt tattaatttg ttcgtatgta ttcaaatata tcctcctcac tattttgatt
     5041 agtacctatt ttatatccat agttgttaat taaataaact taatttagtt tatttataga
     5101 tttcattggc ttctaaattt tttatctaga taataattat tttagttaat tttattctag
     5161 attatatatg atatgatctt tcatttccat aaaactaaag taagtgtaaa cctattcatt
     5221 gttttaaaaa tatctcttgc cagtcacgtt acgttattag ttatagttat tataacatgt
     5281 attcacgaac gaaaatcgcc attcgccagc tgcaggaatt cgacctctag cttgaggcat
     5341 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     5401 gtgaacgctc tcctgagtag gacaaatccg ccgctctagc taagcagaag gccatcctga
     5461 cggatggcct ttttgcgttt ctacaaactc ttgttaactc tagagctgcc tgccgcgttt
     5521 cggtgatgaa gatcttcccg atgattaatt aattcagaac gctcggttgc cgcggcaaga
     5581 acgttgctcg agattggatc caatcatttt gcaaaagttg ttgactttat ctacaaggtg
     5641 tggcatattg gatccaatag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag
     5701 gacgaaacag cctctacaaa taattttgtt taatcggctg taaaggagga attcaaaatg
     5761 acacatgaac attcagcaca atggttgaat aattacaaaa aaggatatgg ttacggtcct
     5821 tatccattag gtataaatgg cggtatgcac tacggagttg atttttttat gaatattgga
     5881 acaccagtaa aagctatttc aagcggaaaa atagttgaag ctggttggag taattacgga
     5941 ggaggtaatc aaataggtct tattgaaaat gatggagtgc atagacaatg gtatatgcat
     6001 ctaagtaaat ataatgttaa agtaggagat tatgtcaaag ctggtcaaat aatcggttgg
     6061 tctggaagca ctggttattc tacagcacca catttacact tccaaagaat ggttaattca
     6121 ttttcaaatt caactgccca agatccaatg cctttcttaa agagcgcagg atatggaaaa
     6181 gcaggtggta cagtaactcc aacgccgaat acaggttgga aaacaaacaa atatggcaca
     6241 ctatataaat cagagtcagc tagcttcaca cctaatacag atataataac aagaacgact
     6301 ggtccattta gaagcatgcc gcagtcagga gtcttaaaag caggtcaaac aattcattat
     6361 gatgaagtga tgaaacaaga cggtcatgtt tgggtaggtt atacaggtaa cagtggccaa
     6421 cgtatttact tgcctgtaag aacatggaat aaatctacta atactttagg tgttctttgg
     6481 ggaactataa agtaacgtcg tgactgggaa aaccctggcg actagtcttg gactcctgtt
     6541 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     6601 ccgggcgttt tttattggtg agaatccagg ggtccccaat aattacgatt taaattggag
     6661 cttagtcgac agctagccgc atgcaagcta attcggtgga aacgaggtca tcatttcctt
     6721 ccgaaaaaac ggttgcattt aaatcttaca tatgtaatac tttcaaagac tacatttgta
     6781 agatttgatg tttgtagcgc ccgtcggctg aaaggtggtg aactactatg gacatgcctc
     6841 gtattaaacc gggtcagcgt gttatgatgg cactgcgtaa aatgattgca agcggtgaaa
     6901 tcaaaagtgg tgaacgtatt gcagaaattc cgaccgcagc agcactgggt gttagccgta
     6961 tgccggttcg tatcgcactg cgttcactgg aacaagaagg tctggttgtt cgtctgggtg
     7021 cacgtggtta tgcagcccgt ggtgttagca gcgatcagat tcgtgatgca attgaagttc
     7081 gtggtgttct ggaaggtttt gcagcacgtc gtctggcaga acgtggtatg accgcagaaa
     7141 cccatgcacg ttttgttgta ctgattgcag aaggtgaagc actgtttgca gccggtcgcc
     7201 tgaatggtga agatctggat cgttatgccg catataatca ggcatttcat gataccctgg
     7261 ttagcgcagc aggtaatggt gcagttgaaa gcgcactggc acgtaatggt tttgaaccgt
     7321 ttgcagcagc cggtgcactg gccctggatc tgatggacct gtctgccgaa tatgaacatc
     7381 tgctggcagc acatcgtcag catcaggcag ttctggatgc agttagctgt ggtgatgccg
     7441 aaggtgcaga acgtattatg cgtgatcatg cactggcagc aattcgtaat gcaaaagttt
     7501 ttgaagcagc agcaagcgca ggcgcaccgc tgggtgcagc atggtcaatt cgtgcagatt
     7561 gataaggatc ctaattggta acgaatcaga caattgacgg cgcgcaacgc aattaatgtg
     7621 agttaggatc ctgagcgccg gtcgctacca ttaccagttg gtctggtgtc aaaaataata
     7681 ataaccgggc aggccatgtc tgcccgtatt tcgcgtaagg aaatccatta tgtactattt
     7741 aattctgcgt gacatcccat cgatcagacc agtttttaat ttgtgtgttt ccatgtgtcc
     7801 agtttggaat actcttaacc tcattggaaa tcgcggcata atcactggtg gtatgattga
     7861 tgaccgcgtc aacaatgacc tttatgccat attcttcagc ggctgcacac atttctttaa
     7921 attcttgttc agtacctaag taacggttgc caatttgata cgatgtcggc tgatacagcc
     7981 agtaccagtt cgacatgctt ttatctcctt gattcccttc ctttacttgg ttaatcggag
     8041 atgtctgaat ggctgtatat cctgcatcat gaatatcctt catattgtgt tttaacgtat
     8101 tgaacgacca attccatgca tgaagaatgg ttccgctttt gatcgacggt gctgtaagct
     8161 cattcgattt gttcgccgtt tcagcactcg cagccgccgg tcctgccaga accaaatgaa
     8221 acagcaataa aaatccagcg aataacggca gtaaagaggt tttgaatcgt tttgcaaaca
     8281 ttcttgacac tccttatttg attttttgaa gacttacttc ggagtcaaaa atccctctta
     8341 cttcattctt ccgcttcctc ctttcaaacc gatgtgaaga ctggagaatt ttgtt
//
