LOCUS       V002806                 8365 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002806
VERSION     V002806
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8365)
  AUTHORS   Hirayama Y, Sakanaka M, Fukuma H, Murayama H, Kano Y, Fukiya S,
            Yokota A.
  TITLE     Development of a Double-Crossover Markerless Gene Deletion System in
            Bifidobacterium longum: Functional Analysis of the
            alpha-Galactosidase Gene for Raffinose Assimilation
  JOURNAL   Appl. Environ. Microbiol. 78 (14), 4984-4994 (2012)
   PUBMED   22582061
REFERENCE   2  (bases 1 to 8365)
  AUTHORS   Hirayama Y, Sakanaka M, Fukuma H, Murayama H, Kano Y, Fukiya S,
            Yokota A.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2011) Contact:Satoru Fukiya Hokkaido University,
            Laboratory of Microbial Physiology, Research Faculty of Agriculture;
            Kita 9, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan
REFERENCE   3  (bases 1 to 8365)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8365)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2012"; volume: "78"; issue: "14";
            pages: "4984-4994"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (11-DEC-2011) Contact:Satoru Fukiya Hokkaido University, Laboratory
            of Microbial Physiology, Research Faculty of Agriculture; Kita 9,
            Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(109..756)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    1046..1393
                     /label="derived from Escherichia coli plasmid pBR322"
                     /note="derived from Escherichia coli plasmid pBR322"
     misc_feature    1104..1393
                     /gene="tetA"
                     /note="tetA was disrupted by the insertion of
                     Bifidobacterium longum BK25 plasmid pTB4"
     gene            1104..1393
                     /gene="tetA"
                     /label="tetA"
     misc_feature    1394..4351
                     /note="derived from Bifidobacterium longum BK25 plasmid
                     pTB4"
     CDS             complement(1689..2249)
                     /codon_start=1
                     /gene="pTB4_p1"
                     /product="hypothetical protein"
                     /label="pTB4_p1"
                     /protein_id="BAL42613.1"
                     /translation="MTNQLLTPAQCPAHRRRIRDAVPGRVQAGHRAQRLGAGTGVRAPL
                     PLLCGRALPAHGREHPRQRDDAAQGAQGEGRWKMTFQKKEEPLPVRGRRHHPERSSTLM
                     RFPMRSDGYIHGHPHGHPHGHLHGHLHGHLHGHLHGHLHGHPHGDVHQIEGKQARRFMY
                     MNRARSLPRPSAQEQQPECRRPE"
     gene            complement(1689..2249)
                     /gene="pTB4_p1"
                     /label="pTB4_p1"
     CDS             2402..3325
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication initiation protein RepA"
                     /label="repA"
                     /protein_id="BAL42614.1"
                     /translation="MSNEYVQGTLELTRAFDGWWLPQRPLCCDDDYSQLVRRSRIDALR
                     CKHIEANPAAQVNMLVVDVDSSEGRLMSLWGHRDMPPNFIAENPANGHCHAGWVLTEPV
                     CRTDMARLKPLKLLHAVTEGLRRSVDGDEGYSGLLMKNPLSDAWDSDLCREDTYDLPDL
                     VAALEAHGDMPPKSWTRTKRAREVGVGRNCTLFDEARTLAYREVRRLPDRTPASSDLLR
                     EYVRRTCHEINASFPDPLPVREVNDTAKSIHKWITTRSRMWRDGAVANAATFVAIQSAR
                     GKKSGEARQNAFEEKFAQYAQEVLGQ"
     gene            2402..3325
                     /gene="repA"
                     /label="repA"
     CDS             3322..3612
                     /codon_start=1
                     /gene="repB"
                     /product="plasmid replication initiation protein RepB"
                     /label="repB"
                     /protein_id="BAL42615.1"
                     /translation="MTIQTIRKKRPLPAKELAEAYGVSVRTIKYWNSQTREDWIDEQAT
                     LRESIRAYHDDDGHSWSQTAEHFNMTQGAVRQRAYRARKEREAEAKAARPE"
     gene            3322..3612
                     /gene="repB"
                     /label="repB"
     CDS             complement(3813..4094)
                     /codon_start=1
                     /gene="pTB4_p2"
                     /product="hypothetical protein"
                     /label="pTB4_p2"
                     /note="Similar to Blon_0217 (Ribbon-helix-helix protein,
                     copG family) [Bifidobacterium longum subsp. infantis ATCC
                     15697]"
                     /protein_id="BAL42616.1"
                     /translation="MSDFSERDRQLMALVGLTEEQVLEDERMAESETVPDDLTGRVYYG
                     LHLAEPDEQMVSVSVRMPKSTLDGLTAMARRYHISRSEYMRRKLAGAI"
     gene            complement(3813..4094)
                     /gene="pTB4_p2"
                     /label="pTB4_p2"
     CDS             complement(4091..4333)
                     /codon_start=1
                     /gene="pTB4_p3"
                     /product="hypothetical protein"
                     /label="pTB4_p3"
                     /protein_id="BAL42617.1"
                     /translation="MERHPELSEKDVVTAFRSVMVDAERESGAWMAIGLDGRGRNVEML
                     YRVVGDLVVIYHAFTPPTKKFRREIDRLRGDGRTL"
     gene            complement(4091..4333)
                     /gene="pTB4_p3"
                     /label="pTB4_p3"
     misc_feature    4352..5252
                     /gene="tetA"
                     /note="tetA was disrupted by the insertion of
                     Bifidobacterium longum BK25 plasmid pTB4"
     gene            4352..5252
                     /gene="tetA"
                     /label="tetA"
     CDS             5891..6079
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    6184..6324
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6510..7098)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7272..8129)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8130..8234)
                     /label="AmpR promoter"
ORIGIN
        1 aagcttggat ctggagctgt aatataaaaa ccttcttcaa ctaacggggc aggttagtga
       61 cattagaaaa ccgactgtaa aaagtacagt cggcattatc tcatattata aaagccagtc
      121 attaggccta tctgacaatt cctgaataga gttcataaac aatcctgcat gataaccatc
      181 acaaacagaa tgatgtacct gtaaagatag cggtaaatat attgaattac ctttattaat
      241 gaattttcct gctgtaataa tgggtagaag gtaattacta ttattattga tatttaagtt
      301 aaacccagta aatgaagtcc atggaataat agaaagagaa aaagcatttt caggtatagg
      361 tgttttggga aacaatttcc ccgaaccatt atatttctct acatcagaaa ggtataaatc
      421 ataaaactct ttgaagtcat tctttacagg agtccaaata ccagagaatg ttttagatac
      481 accatcaaaa attgtataaa gtggctctaa cttatcccaa taacctaact ctccgtcgct
      541 attgtaacca gttctaaaag ctgtatttga gtttatcacc cttgtcacta agaaaataaa
      601 tgcagggtaa aatttatatc cttcttgttt tatgtttcgg tataaaacac taatatcaat
      661 ttctgtggtt atactaaaag tcgtttgttg gttcaaataa tgattaaata tctcttttct
      721 cttccaattg tctaaatcaa ttttattaaa gttcatttga tatgcctcct aaatttttat
      781 ctaaagtgaa tttaggaggc ttacttgtct gctttcttca ttagaatcaa tcctttttta
      841 aaagtcaata ttactgtaac ataaatatat attttaaaaa tatcccactt tatccaattt
      901 tcgtttgttg aactaatggg tgctttagtt gaagaataaa agaccacatt aaaaaatgtg
      961 gtcttttgtg tttttttaaa ggatttgagc gtagcgaaaa atccttttct ttcttatctt
     1021 gataataagg gtaactattg ccgcaagctt gttaatgcgg tagtttatca cagttaaatt
     1081 gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca
     1141 ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc
     1201 gggatatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg
     1261 cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc
     1321 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
     1381 cacccgtcct gtggatcctc atcagcacca cccgcgacac gaacgtatta catacgcaat
     1441 catatcagtg tcacacatca ggtacaaccg taccgcaaca cgcttcgagc ggacgcaagc
     1501 ggcttttgcg cggagcattt ccggccattt gcccagcttg ttccgcgctt ccgagaaggt
     1561 gtggagttgg gttttctgtg atgccgcgcg agcggtctgg aggctccgcc gaccatgggg
     1621 catgggtctt cccggcaagc cgattcgcac cgcgaatcga gtgattgtag ctacaatcac
     1681 gtcgcttgct actccggcct tcggcactcg ggttgttgct cctgcgccga agggcgaggg
     1741 aggctgcggg ctcggttcat gtacatgaac cgtctagctt gcttaccttc gatttgatgg
     1801 acatctccat gtggatgtcc atggagatgt ccatggagat gtccatggag atgtccatgg
     1861 agatgtccat ggagatgtcc atgtgggtgt ccatgtgggt gtccatggat atatccatct
     1921 gagcgcatag gaaaacgcat gagggtggat gatcgttcgg ggtgatgcct tcttccacga
     1981 acgggaaggg gctcttcctt tttctggaat gtcatcttcc agcgcccttc gccttgagct
     2041 ccttgagcag catcgtcgcg ttgtcgcgga tgctcgcgtc cgtgagcggg gagagctcgt
     2101 ccgcatagga gcggcagagg agcgcgtaca cccgttcccg cacccaggcg ctgcgcgcga
     2161 tgcccagctt gaacgcggcc cggcacagca tctcgtattc gtcgccggtg agcaggacat
     2221 tgagctggcg tcaaaagttg attagtcatg ccgtgtatat acgtccgggg tgttttcggc
     2281 cagattccaa cagttttggg ccactccggg aaggaacagc cgacgcttcg ctgggctgaa
     2341 ccatctgact tggttcggcg cacgcttgtg caatgcacaa atgtgcaata ggatgtgagc
     2401 tatgagcaac gaatatgtgc aggggacgct ggagctgacg cgggcgttcg acggctggtg
     2461 gcttccccag cgtcccctgt gctgcgacga tgactactcc cagctggtgc gccggagccg
     2521 catcgacgcg ctcagatgca agcacatcga ggcgaatccg gcagcgcagg tgaacatgct
     2581 cgttgttgac gttgattcgt ctgagggccg tttaatgagc ctgtgggggc atcgggacat
     2641 gccgcccaac ttcatcgcgg agaatccggc caacggccac tgccacgcgg gctgggtgct
     2701 caccgaaccc gtctgccgca ccgacatggc cagactcaag cccctgaagc tcctgcacgc
     2761 cgtcaccgag gggctgcgcc gcagcgtgga cggcgacgag ggctattccg gcctgctcat
     2821 gaagaacccg ttgagcgacg cgtgggacag cgacctgtgc cgcgaggaca cctacgacct
     2881 gcccgacctc gtggccgcgc tcgaagcgca cggggacatg ccgcccaaga gctggacgcg
     2941 caccaaacgc gcccgcgagg tcggagtggg ccgcaactgc accctgttcg acgaggcccg
     3001 caccctcgcc taccgcgagg tgcgccgact gcctgaccgc acgcccgcct cctccgacct
     3061 gctgcgcgag tacgtgcgcc gcacctgcca cgaaatcaac gcctcgttcc ccgacccgct
     3121 gcccgtgcgc gaggtcaacg acaccgccaa gagcatccac aagtggatca ccacgcgaag
     3181 ccgcatgtgg cgagacggtg ccgtcgccaa cgccgccacg ttcgtcgcca tccaatccgc
     3241 acgcggaaag aaatcgggcg aggccagaca aaacgcattt gaagagaagt tcgcgcagta
     3301 cgcgcaggag gtgcttggac aatgaccatc cagacaattc gcaagaagcg cccattgccc
     3361 gccaaggagt tagccgaggc ctatggggtg tccgtgcgca cgatcaagta ctggaattcg
     3421 caaacgcgcg aggactggat agacgagcag gcaacgttgc gcgaatccat ccgcgcctac
     3481 cacgacgatg acggccactc gtggagccag accgccgaac acttcaacat gacccaaggt
     3541 gccgtgcgcc aacgcgccta ccgcgcccgc aaggaacgcg aggccgaggc aaaagccgca
     3601 aggccggagt agcaagtgac gtgaacgtat atacgttcac tcgattcgcg gtgcgaatcg
     3661 gcttgccggg aagacccatg ccccatagtc ggcagagcct ccagaccgct cacgcggcat
     3721 cgtgccacga ctgtggcaca tcacgtaaca cccgcacatc ctcacggtgg tgcgcgggtg
     3781 ttacgtgatt cgaaccatcg gcctgacagg tctcagatcg ctccagcgag cttacggcgc
     3841 atgtactcgc tgcggctgat gtggtaccgc ctggccatcg cggtcagacc gtccagcgtg
     3901 ctcttgggca tgcggacgga cacgctgacc atctgctcgt cgggctcggc gagatggagc
     3961 ccgtagtaca cgcgccccgt caggtcgtcc gggacagtct cggactcggc catccgctca
     4021 tcctccagca cctgctcctc ggtcagcccg accagcgcca tcaactgcct gtcgcgttcg
     4081 ctgaaatcac tcatagcgtc ctcccatcgc cgcgcagacg gtctatctcg cgcctgaact
     4141 tcttggtcgg cggcgtgaac gcgtggtaga tcaccacgag gtcgccgacg accctgtaca
     4201 gcatctcgac gttcctgccg cgcccgtcga gccctatcgc catccacgcg ccggactcgc
     4261 gctccgcgtc caccatgacg ctgcggaacg cggtcaccac gtccttctcg gacagttctg
     4321 ggtgacgctc catcaccctg tcaagcacct ggatcctcta cgccggacgc atcgtggccg
     4381 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     4441 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     4501 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     4561 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     4621 gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     4681 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4741 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     4801 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     4861 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     4921 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     4981 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     5041 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     5101 cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     5161 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     5221 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg
     5281 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac
     5341 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat
     5401 ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac
     5461 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
     5521 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
     5581 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
     5641 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
     5701 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
     5761 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
     5821 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac
     5881 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
     5941 ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat
     6001 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
     6061 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6181 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     6241 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     6301 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6361 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6421 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6481 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6541 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6601 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6661 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6721 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6781 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6841 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6901 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6961 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7021 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7081 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7141 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7201 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7261 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7321 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7381 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7441 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7501 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7561 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     7621 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7681 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7741 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7801 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7861 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     7921 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7981 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8041 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8101 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8161 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8221 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     8281 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc
     8341 tcatgtttga cagcttatca tcgat
//
