LOCUS       V005221                 6921 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005221
VERSION     V005221
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6921)
  AUTHORS   Sakka K, Wang YY, Sakka M, Kunitake E, Kimura T.
  TITLE     A plasmid vector for Ruminiclostridium josui
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6921)
  AUTHORS   Sakka K, Kimura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-OCT-2017) Contact:Kazuo Sakka Mie University, Graduate
            School of Bioresources; 1577 Kurimamachiya-cho, Tsu, Mie 514-8507,
            Japan URL :http://www.bio.mie-u.ac.jp/
REFERENCE   3  (bases 1 to 6921)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6921)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-OCT-2017) Contact:Kazuo Sakka Mie University, Graduate School of
            Bioresources"; volume: " 1577 Kurimamachiya-cho, Tsu, Mie 514-8507,
            Japan URL :http"; pages: "//www.bio.mie-u.ac.jp"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..194
                     /label="AmpR promoter"
     CDS             195..1052
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1226..1814
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2102..2123
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2138..2168
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2176..2192
                     /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)."
     primer_bind     2200..2216
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      2232..2275
                     /label="pcip promoter"
                     /note="pcip promoter"
                     /regulatory_class="promoter"
     RBS             2558..2566
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2824..2841
                     /label="6xHis"
                     /note="6xHis affinity tag"
     primer_bind     complement(2854..2870)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3458..3687
                     /note="oriCP; C. perfringens pIP404 replication origin"
     CDS             complement(3750..4970)
                     /codon_start=1
                     /gene="rep"
                     /product="pIP404 replication"
                     /label="rep"
                     /protein_id="BBA94093.1"
                     /translation="MRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAELK
                     DLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYFNQ
                     SIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDATRIL
                     RVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYRERSL
                     YYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSMFRQHL
                     SEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEYKRRENI
                     RGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKTLERHITY
                     MKKNGLL"
     gene            complement(3750..4970)
                     /gene="rep"
                     /label="rep"
     CDS             complement(6032..6766)
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
ORIGIN
        1 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
       61 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      421 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
      601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
      661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
      721 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
      781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
      841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
      901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2041 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2101 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     2161 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     2221 attacgaatt catattcatt tcaaatatct gtaagactcg ctttataagt attaattctc
     2281 taagttatat tctttctaac tatcattata tctaaattta aattacgtct acgaactatt
     2341 tggctatatt tagctacaag cattatgaca gaatagtacc ttcttttcag aaaaagttat
     2401 cgttatttaa aaaaatataa taaaagtgat tataaaaagt atcaaaataa tcggaataga
     2461 tgactgtggt aatgtgtata ttttgagaat tttataatca agttaaagca tttctgaatt
     2521 aatttaaatt atgaaaaacc acttagccaa taaagattaa ggaggtgaaa atatttattc
     2581 agttatatcg gtgtgcctgt aaaaatgaaa cattttaaaa catttttaga gtaaggcagt
     2641 ttaaaagcat cttttattta aaagtacttt ttattaataa ctgccttaaa ctgataaaag
     2701 actgaatatt ggtgataaaa cccatagtca aaaggccaga tggttttcat gtaaaataaa
     2761 aaataggagg tttacccatg gcggggatcc tctagagtcg acctgcaggc ggccgcactc
     2821 gagcaccacc accaccacca ctgaaagctt ggcactggcc gtcgttttac aacgtcgtga
     2881 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2941 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3001 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3061 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3121 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3181 acaagctgtg accgtctccg ggaccaataa tcccactcta agccacaaac acgccctata
     3241 aaatcccgct ttaatcccac tttgagacac atgtaatatt actttacgcc ctagtatagt
     3301 gataattttt tacattcaat gccacgcaaa aaaataaagg ggcactataa taaaagttcc
     3361 ttcggaacta actaaagtaa aaaattatct ttacaacctc cccaaaaaaa agaacaggta
     3421 caaagtaccc tataatacaa gcgtaaaaaa atgagggtaa aaataaaaaa ataaaaaaat
     3481 aaaaaaataa aaaaataaaa aaaataaaaa aataaaaaaa taaaaaaata aaaaaataaa
     3541 aaaataaaaa aataaaaaaa taaaaaaata taaaaataaa aaaatataaa aataaaaaaa
     3601 tataaaaata aaaaaatata aaaataaaaa aataaaaaaa tataaaaata aaaaaataaa
     3661 aaaatataaa aatatttttt atttaaagtt tgaaaaaaat ttttttatat tatataatct
     3721 ttgaagaaaa gaatataaaa aatgagcctt tataaaagcc catttttttt catatacgta
     3781 atatgacgtt ctaatgtttt tattggtact tctaacatta gagtaatttc tttattttta
     3841 aagccttttt ctttaagggc ttttattttt tttcttaata catttaattc ctcttttttt
     3901 gttgcttttc ctttagcttt taattgctct tgataatttt ttttacctct aatattttct
     3961 cttctcttat attccttttt agaaattatt attgtcatat atttttgttc ttcttctgta
     4021 atttctaata actctataag agtttcattc ttatacttat attgcttatt tttatctaaa
     4081 taacatcttt cagcacttct agttgctctt ataacttctc tttcacttaa atgttgtcta
     4141 aacatactat taagttctaa aacatcattt aatgccttct caatgtcttc tgtaaagcta
     4201 caaagataat atctatataa aaataatata agctctctgt gtccttttaa atcatattct
     4261 cttagttcac aaagttttat tatgtcttgt attcttccat aatataaact tctttctcta
     4321 taaatataat ttattttgct tggtctaccc tttttccttt catatggttt taattcaggt
     4381 aaaaatccat tttgtatttc tcttaagtca taaatatatt cgtactcatc taatatattg
     4441 actactgttt ttgatttaga gtttatactt cctggaactc ttaatattct ggttgcatct
     4501 aaggcttgtc tatctgctcc aaagtatttt aattgattat ataaatattc ttgaaccgct
     4561 ttccataatg gtaatgcttt actaggtact gcatttatta tccatattaa atacattcct
     4621 cttccactat ctattacata gtttggtata ggaatacttt gattaaaata attcttttct
     4681 aagtccatta atacctggtc tttagttttg ccagttttat aataatccaa gtctataaac
     4741 agtgtattta actcttttat attttctaat cgcctacacg gcttataaaa ggtatttaga
     4801 gttatataga tattttcatc actcatatct aaatctttta attcagcgta tttatagtgc
     4861 cattggctat atcctttttt atctataacg ctcctggtta tccacccttt acttctacta
     4921 tgaatattat ctatatagtt ctttttattc agctttaatg cgtttctcac ttattcacct
     4981 ccccttctgt aaaactaaga aaattatatc atattttcaa taattattaa ctattcttaa
     5041 actcttaata aaaaatagag taagtcccca attgaaactt aatctatttt ttatgtttta
     5101 atttattatt tttattaaaa tattttaaac taaattaaat gattcttttt aattttttac
     5161 tatttcattc cataatatat tactataatt atttacaaat aatatttctt catttgtaat
     5221 atttagatga tttactaatt ttagttttta tatattaaat aattaatgta taatttatat
     5281 aaaaaatcaa aggagcttat aaattatgat tatttccaaa gatactaaag atttaatttt
     5341 ttcaatttta acaatacttt ttgtaatatt atgtttaaat ttaattgtat ttttttcata
     5401 taataaagcc gttgaagtaa accaatccat tttccttatg atgttattat taaatttaag
     5461 ttttataata atatctttat tatatttatt gtttttaaaa aaactagtga aatttccggc
     5521 tttattaaac ttatttttag gaattttatt ttcattttca tctttacagg atttgattat
     5581 atctttaaat atgttttatc aaatattatc tttttctaaa tttatatata tttttattat
     5641 atttattatt atatatattt tatttttaag tttctttcta acagctatta aaaagaaact
     5701 taaaaataaa aacacgtact ctaaaccaat aaataaaact atttttatta ttgctgcctt
     5761 gattggaata gtttttagta aaattaattt caatattcca caatattata ttataagcta
     5821 gctttgcatt gtacttttca atcgcttcac gaatgcggtt atctccgaaa gataaagtct
     5881 tttcatcttc cttgatgaag ataagatttt ctccgtctcc gccgacctct ttagctcctt
     5941 ggaagctgtc agtagtatac ctaataattt atctacattc cctttagtaa cgtgtaactt
     6001 tccaaattta caaaagcgac tcatagaatt atttcctccc gttaaataat agataactat
     6061 taaaaataga caatacttgc tcataagtaa cggtacttaa attgtttact ttggcgtgtt
     6121 tcattgcttg atgaaactga tttttagtaa acagttgacg atattctcga ttgacccatt
     6181 ttgaaacaaa gtacgtatat agcttccaat atttatctgg aacatctgtg gtatggcggg
     6241 taagttttat taagacactg tttacttttg gtttaggatg aaagcattcc gctggcagct
     6301 taagcaattg ctgaatcgag acttgagtgt gcaagagcaa ccctagtgtt cggtgaatat
     6361 ccaaggtacg cttgtagaat ccttcttcaa caatcagata gatgtcagac gcatggcttt
     6421 caaaaaccac ttttttaata atttgtgtgc ttaaatggta aggaatactc ccaacaattt
     6481 tatacctctg tttgttaggg aattgaaact gtagaatatc ttggtgaatt aaagtgacac
     6541 gagtattcag ttttaatttt tctgacgata agttgaatag atgactgtct aattcaatag
     6601 acgttacctg tttacttatt ttagccagtt tcgtcgttaa atgcccttta cctgttccaa
     6661 tttcgtaaac ggtatcggtt tcttttaaat tcaattgttt tattatttgg ttgagtactt
     6721 tttcactcgt taaaaagttt tgagaatatt ttatattttt gttcatgtaa tcactccttc
     6781 ttaattacaa atttttagca tctaatttaa cttcaattcc tattatacaa aattttaaga
     6841 tactgcacta tcaacacact cttaagtttg cttctaagtc ttatttccat aacttctttt
     6901 acgtttccgg gtacaattcg t
//
