LOCUS       V016242                 7446 bp    DNA     circular SYN 02-SEP-2021
DEFINITION  Exported.
ACCESSION   V016242
VERSION     V016242
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7446)
  AUTHORS   Suzuki S, Osada S, Imamura D, Sato T.
  TITLE     New Bacillus subtilis vector, pSSbeta as genetic tool for
            site-specific integration and excision of cloned DNA, and prophage
            elimination
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7446)
  AUTHORS   Suzuki S, Sato T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2021) Contact:Shota Suzuki Department of Life
            Science, College of Science, Rikkyo University; 3-34-1
            Nishi-Ikebukuro, Toshima-ku, Tokyo 171-8501, Japan
REFERENCE   3  (bases 1 to 7446)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (26-AUG-2021) Contact:Shota Suzuki Department of Life Science,
            College of Science, Rikkyo University; 3-34-1 Nishi-Ikebukuro,
            Toshima-ku, Tokyo 171-8501, Japan"
FEATURES             Location/Qualifiers
     source          1..7446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            361..1998
                     /gene="sprA"
                     /label="sprA"
     CDS             361..1998
                     /codon_start=1
                     /transl_table=11
                     /gene="sprA"
                     /product="serine-type phage integrase"
                     /label="sprA"
                     /note="phage Spbeta"
                     /protein_id="BDB04359.1"
                     /translation="MELKNIVNSYNITNILGYLRRSRQDMEREKRTGEDTLTEQKELMN
                     KILTAIEIPYELKMEIGSGESIDGRPVFKECLKDLEEGKYQAIAVKEITRLSRGSYSDA
                     GQIVNLLQSKRLIIITPYKVYDPRNPVDMRQIRFELFMAREEFEMTRERMTGAKYTYAA
                     QGKWISGLAPYGYQLNKKTSKLDPVEDEAKVVQLIFNIFLNGLNGKDYSYTAIASHLTN
                     LQIPTPSGKKRWNQYTIKAILQNEVYIGTVKYKVREKTKDGKRTIRPEKEQIVVQDAHA
                     PIIDKEQFQQSQVKIANKVPLLPNKDEFELSELAGVCTCSKCGEPLSKYESKRIRKNKD
                     GTESVYHVKSLTCKKNKCTYVRYNDVENAILDYLSSLNDLNDSTLTKHINSMLSKYEDD
                     NSNMKTKKQMSEHLSQKEKELKNKENFIFDKYESGIYSDELFLKRKAALDEEFKELQNA
                     KNELNGLQDTQSEIDSNTVRNNINKIIDQYHIESSSEKKNELLRMVLKDVIVNMTQKRK
                     GPIPAQFEITPILRFNFIFDLTATNSFH"
     gene            2023..2038
                     /gene="attP"
                     /label="attP"
     misc_feature    2023..2038
                     /gene="attP"
                     /label="Bacteriophage SPbeta attachment site"
                     /note="Bacteriophage SPbeta attachment site"
     gene            complement(2186..2362)
                     /gene="sprB"
                     /label="sprB"
     CDS             complement(2186..2362)
                     /codon_start=1
                     /transl_table=11
                     /gene="sprB"
                     /product="recombination directionality factor"
                     /label="sprB"
                     /note="phage Spbeta"
                     /protein_id="BDB04360.1"
                     /translation="MEPYQRYEELKKKTIKVVQKENYSIRYITQDEASNDLDEFYKQFA
                     QHLLEAALERKAE"
     protein_bind    2442..2466
                     /label="xyl repressor encoded by xylR binding site"
                     /bound_moiety="xyl repressor encoded by xylR"
     regulatory      2476..2481
                     /label="Pxyl_-10"
                     /note="Pxyl_-10"
                     /regulatory_class="minus_10_signal"
     regulatory      2498..2504
                     /label="Pxyl_-35"
                     /note="Pxyl_-35"
                     /regulatory_class="minus_35_signal"
     gene            2610..3764
                     /gene="xylR"
                     /label="xylR"
     CDS             2610..3764
                     /codon_start=1
                     /transl_table=11
                     /gene="xylR"
                     /product="transcriptional regulator"
                     /label="xylR"
                     /protein_id="BDB04361.1"
                     /translation="MDIADQTFVKKVNQKLLLKEILKNSPISRAKLSEMTGLNKSTVSS
                     QVNTLMKESMVFEIGQGQSSGGRRPVMLVFNKKAGYSVGIDVGVDYINGILTDLEGTIV
                     LDQYRHLESNSPEITKDILIDMIHHFITQMPQSPYGFIGIGICVPGLIDKDQKIVFTPN
                     SNWRDIDLKSSIQEKYNVSVFIENEANAGAYGEKLFGAAKNHDNIIYVSISTGIGIGVI
                     INNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRGCWELYASEKALLKSLQTKEKKLSYQ
                     DIINLAHLNDIGTLNALQNFGFYLGIGLTNILNTFNPQAVILRNSIIESHPMVLNSMRS
                     EVSSRVYSQLGNSYELLPSSLGQNAPALGMSSIVIDHFLDMITM"
     CDS             complement(4065..4712)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     primer_bind     5139..5155
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    5156..5212
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5225..5241)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5249..5265)
                     /label="lac operator"
                     /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)."
     promoter        complement(5273..5303)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5318..5339)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5627..6215)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6389..7246)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7247..7351)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca ctttcagcac acccaccagt gaagaaaata
      121 aaagatacac aaactaacat tgaaaagcga atattcataa agccctcctg gatttataaa
      181 gtatttacaa tgaatttaga tgaactaatt ataacccatg ttcaagaagg tttttcataa
      241 aatttcaatc aattcaattc cttcaaatta ttgtattgtt ttcgtattca atgtcagata
      301 aaatattctt aattaagttt ctgttttgac attaaacaca aataaagagg tgctaaattt
      361 ttggagttaa aaaacattgt taattcttac aacatcacaa atattttagg gtatcttaga
      421 cgttctcgtc aggacatgga acgagaaaaa agaacaggtg aagacacact cacagaacaa
      481 aaagaactta tgaataaaat acttacggcc attgagatcc cttatgaatt aaaaatggag
      541 attggatcag gagaaagcat tgatggtcga cctgttttca aagaatgtct taaagatctt
      601 gaggaaggca aatatcaggc aattgctgtt aaagaaataa caagacttag tcgtgggagt
      661 tatagcgatg caggacaaat tgttaacctc cttcaaagca aacgattgat cataattaca
      721 ccatataagg tttatgaccc tagaaaccct gttgacatgc gccagatacg ttttgagtta
      781 tttatggcta gagaagaatt tgaaatgact agggagcgta tgacaggcgc taaatacact
      841 tatgcagcac agggaaaatg gatatctgga ctggcacctt atggatacca attaaacaaa
      901 aagacgtcta aactagatcc agttgaggat gaagcaaaag tagttcaatt aatttttaac
      961 atctttctaa atgggttgaa tggaaaagat tatagctata cagcaattgc ttcacatcta
     1021 acaaatttac aaatccctac accgtcaggg aaaaaacgtt ggaatcaata cacaatcaaa
     1081 gcaattctcc aaaatgaagt ttatatagga actgttaaat ataaggttag agaaaaaaca
     1141 aaagacggaa agagaacaat tagacctgaa aaagaacaaa ttgtagtaca agatgcgcat
     1201 gctcctataa ttgacaaaga acaatttcaa caatcacaag taaaaatcgc aaacaaagtt
     1261 ccccttttgc caaataaaga tgaatttgaa ttaagtgaac tggccggtgt ttgtacttgc
     1321 tcaaaatgtg gtgagccttt atctaaatac gaatctaaac gcatacgaaa aaacaaagat
     1381 ggaactgaaa gcgtttatca cgttaaatcg ctcacatgta agaaaaataa gtgtacgtat
     1441 gtgagatata acgatgttga aaatgcaatt ctggattacc tctcttcctt aaatgactta
     1501 aatgatagta ctttaacaaa acacatcaat tcgatgcttt caaagtacga agacgacaac
     1561 agtaacatga aaacaaaaaa gcaaatgtct gaacaccttt cacaaaaaga gaaagagcta
     1621 aaaaacaaag agaacttcat cttcgataaa tatgaaagtg gcatttacag tgatgaactt
     1681 tttttgaaga gaaaagctgc attagatgaa gaatttaaag agctccagaa tgcaaagaat
     1741 gagctcaatg gattacaaga tacacaatca gaaatagaca gtaatacagt gagaaacaac
     1801 ataaataaaa taattgacca gtatcacatt gaatcaagct cagagaaaaa gaacgagcta
     1861 ttaaggatgg tcttaaaaga tgttattgtt aacatgacac aaaaaagaaa agggcctata
     1921 ccagctcaat ttgaaataac ccctatccta aggtttaatt tcatttttga cctcacagcc
     1981 actaatagtt tccactaaga aaagtagtaa gtatcttaaa aaacagataa agctgtatat
     2041 taagatactt actacatatc taacgaagac aagcccgtca gggcgcgtca gcgggtgttg
     2101 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgcagac caactggtaa
     2161 tggtagcgac cggcgctcac gatgcttact ctgccttcct ttccagagca gcttctaata
     2221 aatgttgagc aaattgttta taaaactcat ctaagtcatt acttgcttca tcctgagtta
     2281 tatatcgaat actgtaattt tctttctgga ctacctttat tgtttttttc tttaattcct
     2341 cataacgttg gtaaggttcc aaggctacac ccccgttttc actatcgatc cggcattcaa
     2401 atacagatgc attttatttc atatagtaag tacatcacct attagtttgt tgtttaaaca
     2461 aactaactta ttttcatctt atataacctc gtcagtattt tcaatatttt ttttagtttt
     2521 ttatgaacac attagattta ataaagggaa gattcgctat gtactatgtt gatacttaat
     2581 ttaaagatta aacaaatgga gtggatgaag tggatatcgc tgatcaaacc tttgtcaaaa
     2641 aagtaaatca aaagttatta ttaaaagaaa tccttaaaaa ttcacctatt tcaagagcaa
     2701 aattatctga aatgactgga ttaaataaat caactgtctc atcacaggta aacacgttaa
     2761 tgaaagaaag tatggtattt gaaataggtc aaggacaatc aagtggcgga agaagacctg
     2821 tcatgcttgt ttttaataaa aaggcaggat actccgttgg aatagatgtt ggtgtggatt
     2881 atattaatgg cattttaaca gaccttgaag gaacaatcgt tcttgatcaa taccgccatt
     2941 tggaatccaa ttctccagaa ataacgaaag acattttgat tgatatgatt catcacttta
     3001 ttacgcaaat gccccaatct ccgtacgggt ttattggtat aggtatttgc gtgcctggac
     3061 tcattgataa agatcaaaaa attgttttca ctccgaactc caactggaga gatattgact
     3121 taaaatcttc gatacaagag aagtacaatg tgtctgtttt tattgaaaat gaggcaaatg
     3181 ctggcgcata tggagaaaaa ctatttggag ctgcaaaaaa tcacgataac attatttacg
     3241 taagtatcag cacaggaata gggatcggtg ttattatcaa caatcattta tatagaggag
     3301 taagcggctt ctctggagaa atgggacata tgacaataga ctttaatggt cctaaatgca
     3361 gttgcggaaa ccgaggatgc tgggaattgt atgcttcaga gaaggcttta ttaaaatctc
     3421 ttcagaccaa agagaaaaaa ctgtcctatc aagatatcat aaacctcgcc catctgaatg
     3481 atatcggaac cttaaatgca ttacaaaatt ttggattcta tttaggaata ggccttacca
     3541 atattctaaa tactttcaac ccacaagccg taattttaag aaatagcata attgaatcgc
     3601 atcctatggt tttaaattca atgagaagtg aagtatcatc aagggtttat tcccaattag
     3661 gcaatagcta tgaattattg ccatcttcct taggacagaa tgcaccggca ttaggaatgt
     3721 cctccattgt gattgatcat tttctggaca tgattacaat gtaatttttt atggaatgga
     3781 cagctcatct ttaaagatga gtttttttat tctaggagta tttctgaagc aatagtgatg
     3841 agctattaag ccgaccattc gacaagtttt gggattgtta agggttccga ggctcaacgt
     3901 caataaagca attggaataa agaagcgaaa aaggagaagt cggttcagaa aaagaaggat
     3961 atggatctgg agctgtaata taaaaacctt cttcaactaa cggggcaggt tagtgacatt
     4021 agaaaaccga ctgtaaaaag tacagtcggc attatctcat attataaaag ccagtcatta
     4081 ggcctatctg acaattcctg aatagagttc ataaacaatc ctgcatgata accatcacaa
     4141 acagaatgat gtacctgtaa agatagcggt aaatatattg aattaccttt attaatgaat
     4201 tttcctgctg taataatggg tagaaggtaa ttactattat tattgatatt taagttaaac
     4261 ccagtaaatg aagtccatgg aataatagaa agagaaaaag cattttcagg tataggtgtt
     4321 ttgggaaaca atttccccga accattatat ttctctacat cagaaaggta taaatcataa
     4381 aactctttga agtcattctt tacaggagtc caaataccag agaatgtttt agatacacca
     4441 tcaaaaattg tataaagtgg ctctaactta tcccaataac ctaactctcc gtcgctattg
     4501 taaccagttc taaaagctgt atttgagttt atcacccttg tcactaagaa aataaatgca
     4561 gggtaaaatt tatatccttc ttgttttatg tttcggtata aaacactaat atcaatttct
     4621 gtggttatac taaaagtcgt ttgttggttc aaataatgat taaatatctc ttttctcttc
     4681 caattgtcta aatcaatttt attaaagttc atttgatatg cctcctaaat ttttatctaa
     4741 agtgaattta ggaggcttac ttgtctgctt tcttcattag aatcaatcct tttttaaaag
     4801 tcaatattac tgtaacataa atatatattt taaaaatatc ccactttatc caattttcgt
     4861 ttgttgaact aatgggtgct ttagttgaag aataaagacc acattaaaaa atgtggtctt
     4921 ttgtgttttt ttaaaggatt tgagcgtagc gaaaaatcct tttctttctt atcttgataa
     4981 taagggtaac tattgccgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5041 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5101 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     5161 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
     5221 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5281 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5821 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6061 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7381 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7441 ttcgtc
//
