LOCUS       sequence-120067-        9840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A transformation-associated recombination (TAR) based genomic 
            capture and expression vector for antibiotic production in B. 
            subtilis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9840)
  AUTHORS   Li Y, Li Z, Yamanaka K, Xu Y, Zhang W, Vlamakis H, Kolter R, Moore 
            BS, Qian PY
  TITLE     Directed natural product biosynthesis gene cluster capture and 
            expression in the model bacterium Bacillus subtilis.
  JOURNAL   Sci Rep. 2015 Mar 24;5:9383. doi: 10.1038/srep09383.
  PUBMED    25807046
REFERENCE   2  (bases 1 to 9840)
  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: "Sci Rep."; 
            date: "2015-03-24"; volume: "5:9383. doi"; pages: " 
            10.1038/srep09383"
COMMENT     Sequence Label: pCAPB02
FEATURES             Location/Qualifiers
     source          1..9840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     2185..2206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2446..3120)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3121..3401)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3418..3437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3537..3559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3656..4159
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     terminator      7309..7395
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7414..7441
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      7521..7555
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    7659..7675
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(7979..7998)
                     /label=LacI-R
                     /note="LacI, reverse primer"
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 gcgtgggtct 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 tatcaactct 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 gtgttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2161 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2221 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2281 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2341 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagatcg
     2401 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     2461 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     2521 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     2581 tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga
     2641 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     2701 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     2761 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     2821 atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg
     2881 ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt
     2941 tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac
     3001 acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct
     3061 ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat
     3121 actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg
     3181 ccctccctct tggccctctc cttttctttt ttctaccgaa ttaattctta atcggcaaaa
     3241 aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa gctatttttc aataaagaat
     3301 atcttccact actgccatct ggcgtcataa ctgcaaagta cacatatatt acgatgctgt
     3361 ctattaaatg cttcctatat tatatatata gtaatgtcgt ttatggtgca ctctcagtac
     3421 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3481 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3541 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3601 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacggatcg
     3661 cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac
     3721 tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta
     3781 gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag
     3841 cgtactttac atatatattt attagacaag aaaagcagat taaatagata tacattcgat
     3901 taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag
     3961 atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg
     4021 ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat
     4081 ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt aaattataat
     4141 tatttttata gcacgtgatg aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg
     4201 aacccctatt tgtttatttt tctaaataca ccatggatca agagacagga tgaggatcgt
     4261 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggtgccctga atgaactgca
     4321 gaaagagctg gtagttggcg cactgttcga agaactgccg atgtccagta agattcttac
     4381 tatgctggtt gaaccggatg ctggtaaagc tacttgggtt gctgcttcta cttatggtac
     4441 cgatacaact actggtgagg aagttaaagg agctcttaaa gaaatccact tcagtaggtg
     4501 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
     4561 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
     4621 agcggtccag tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
     4681 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
     4741 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gactaagaaa
     4801 atgccgtcaa atccgctcgc catgacttca ctaacgatgc ctttgaaaat cttcaagttc
     4861 ttttctacta attcaaggcg tgtctcacca ggtttttggt ttgctccggc gcaaatgcag
     4921 acaatatcag catccttgca gggtatgttt ctctttgatg tctttttgtt tgtgaagtat
     4981 ttcacattta tattgtgcaa cacttcacaa acttttgcaa gagaaaagtt ttgtctgatt
     5041 tatgaacaaa aaagaaacca tcattgatgg tttctttcgg taagtcccgt ctagccttgc
     5101 cctcaatggg gaagagaacc gcttaagccc gagtcattat ataaaccatt tagcacgtaa
     5161 tcaaagccag gctgattctg accgggcact tgggcgctgc cattattaaa aatcactttt
     5221 gcgttggttg tatccgtgtc cgcaggcagc gtcagcgtgt aaattccgtc tgcattttta
     5281 gtcattggtt ttccaggcca agatccggtc aattcaatta ctcggctccc atcatgttta
     5341 tagatataag catttacctg gctccaatga ttcggatttt gatagccgat ggttttggcc
     5401 gacgctggat ctcttttaac aaaactgtat ttctcggtcc tcgttacacc atcactgttc
     5461 gttcctttta acatgatggt gtatgttttg ccaaatggat ctccttttcc gattgtgaat
     5521 tgatctccat ccttaaacgc cgtctctggt ccattattga tttgataaac ggcttttgtt
     5581 gtattcgcat ctgcacgcaa ggtaatcgtc agttgatcat tgaaagaatg tgttacacct
     5641 gttttgtaat tctcaaggaa aacatgaggc gcttttgcaa tatcatcagg ataaagcaca
     5701 gctacagacc tggcattgat cgtgcctgtc agtttaccat cgttcacttg aaatgaaccc
     5761 gctccagctt tattgtcata cctgccatca ggcaattttg ttgccgtatt gatagagaca
     5821 gaggatgaac ctgcatttgc cagcacaacg ccatgtgagc cgcgctgatt cataaatatc
     5881 tggttgtttc cattcgggtt cgagagttcc tcaggctgtc cagccatcac attgtgaaat
     5941 ctattgaccg cagtgatagc ctgatcttca aataaagcac tcccgcgatc gcctatttgg
     6001 cttttccccg ggaacctcac accatttccg cctccctcag gtctggaaaa gaaaagaggc
     6061 gtactgcctg aacgagaagc tatcaccgcc cagcctaaac ggatgatccc cctatgcaag
     6121 ggtttattgt tttctaaaat ctgattacca attagaatga atatttccca aatattaaat
     6181 aataaaacaa aaaaattgaa aaaagtgttt ccaccatttt ttcaattttt ttataatttt
     6241 tttaatctgt tatttaaata gtttatagtt aaatttacat tttcattagt ccattcaata
     6301 ttctctccaa gataactacg aactgctaac aaaattctct ccctatgttc taatggagaa
     6361 gattcagcca ctgcatttcc cgcaatatct tttggtatga ttttacccgt gtccatagtt
     6421 aaaatcatac ggcataaagt taatatagag ttggtttcat catcctgata attatctatt
     6481 aattcctctg acgaatccat aatggctctt ctcacatcag aaaatggaat atcaggtagt
     6541 aattcctcta agtcataatt tccgtatatt cttttatttt ttcgttttgc ttggtaaagc
     6601 attatggtta aatctgaatt taattccttc tgaggaatgt atccttgttc ataaagctct
     6661 tgtaaccatt ctccataaat aaattcttgt ttgggaggat gattccacgg taccatttct
     6721 tgctgaataa taattgttaa ttcaatatat cgtaagttgc ttttatctcc tatttttttt
     6781 gaaataggtc taattttttg tataagtatt tctttacttt gatctgtcaa tggttcagat
     6841 acgacgacta aaaagtcaag atcactattt ggttttagtc cactctcaac tcctgatcca
     6901 aacatgtaag taccaataag gttatttttt aaatgtttcc gaagtatttt tttcacttta
     6961 ttaatttgtt cgtatgtatt caaatatatc ctcctcacta ttttgattag tacctatttt
     7021 atatccatag ttgttaatta aataaactta atttagttta tttatagatt tcattggctt
     7081 ctaaattttt tatctagata ataattattt tagttaattt tattctagat tatatatgat
     7141 atgatctttc atttccataa aactaaagta agtgtaaacc tattcattgt tttaaaaata
     7201 tctcttgcca gtcacgttac gttattagtt atagttatta taacatgtat tcacgaacga
     7261 aaatcgccat tcgccagctg caggaattcg acctctagct tgaggcatca aataaaacga
     7321 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     7381 ctgagtagga caaatccgcc gctctagcta agcagaaggc catcctgacg gatggccttt
     7441 ttgcgtttct acaaactctt gttaactcta gagctgcctg ccgcgtttcg gtgatgaaga
     7501 tcttcccgat gattaattaa ttcagaacgc tcggttgccg ccgggcgttt tttatgcagc
     7561 aatggcaaga acgttgctcg agggtaaatg tgagcactca caattcattt tgcaaaagtt
     7621 gttgacttta tctacaaggt gtggcataat gtgtgtaatt gtgagcggat aacaattaag
     7681 cttagtcgac ccgcggagcg gccgcatcga tgcatgcaag ctaattcggt ggaaacgagg
     7741 tcatcatttc cttccgaaaa aacggttgca tttaaatctt acatatgtaa tactttcaaa
     7801 gactacattt gtaagatttg atgtttgagt cggctgaaag atcgtacgta ccaattattg
     7861 tttcgtgatt gttcaagcca taacactgta gggatagtgg aaagagtgct tcatctggtt
     7921 acgatcaatc aaatattcaa acggagggag acgattttga tgaaaccagt aacgttatac
     7981 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     8041 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     8101 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     8161 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     8221 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     8281 aaaacggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     8341 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     8401 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     8461 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     8521 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggctggcata aatatctcac
     8581 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     8641 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     8701 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     8761 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     8821 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     8881 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     8941 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     9001 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     9061 atgtgagtta ggatcctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa
     9121 taataataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac
     9181 tatttaattc tgcgtgacat cccatcgatc agaccagttt ttaatttgtg tgtttccatg
     9241 tgtccagttt ggaatactct taacctcatt ggaaatcgcg gcataatcac tggtggtatg
     9301 attgatgacc gcgtcaacaa tgacctttat gccatattct tcagcggctg cacacatttc
     9361 tttaaattct tgttcagtac ctaagtaacg gttgccaatt tgatacgatg tcggctgata
     9421 cagccagtac cagttcgaca tgcttttatc tccttgattc ccttccttta cttggttaat
     9481 cggagatgtc tgaatggctg tatatcctgc atcatgaata tccttcatat tgtgttttaa
     9541 cgtattgaac gaccaattcc atgcatgaag aatggttccg cttttgatcg acggtgctgt
     9601 aagctcattc gatttgttcg ccgtttcagc actcgcagcc gccggtcctg ccagaaccaa
     9661 atgaaacagc aataaaaatc cagcgaataa cggcagtaaa gaggttttga atcgttttgc
     9721 aaacattctt gacactcctt atttgatttt ttgaagactt acttcggagt caaaaatccc
     9781 tcttacttca ttcttccgct tcctcctttc aaaccgatgt gaagactgga gaattttgtt
//
