LOCUS       V002051                 6274 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002051
VERSION     V002051
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S, Kuwahara T, Okabe A.
  TITLE     Development and characterization of a xylose-inducible gene
            expression system for Clostridium perfringens
  JOURNAL   Appl. Environ. Microbiol. 77 (23), 8439-8441 (2011)
   PUBMED   21965407
REFERENCE   2  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S.
  TITLE     Development of xylose-inducible EGFP fusion Expression Vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6274)
  AUTHORS   Nariya H, Miyata S.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2018) Contact:Hirofumi Nariya Graduate School of
            Biosphere Science, Hiroshima University, Laboratory of Food
            Microbiology and Hygiene; 1-4-4, Kagamiyama 1-chome,
            Higashi-Hiroshima, Hiroshima 739-8528, Japan
REFERENCE   4  (bases 1 to 6274)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6274)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2011"; volume: "77"; issue: "23";
            pages: "8439-8441"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (07-APR-2018) Contact:Hirofumi Nariya Graduate School of Biosphere
            Science, Hiroshima University, Laboratory of Food Microbiology and
            Hygiene; 1-4-4, Kagamiyama 1-chome, Higashi-Hiroshima, Hiroshima
            739-8528, Japan"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      266..430
                     /note="ori; derived from Clostridium perfringens plasmid
                     pIP404 (INSD:M32882.1)"
     CDS             complement(493..1713)
                     /codon_start=1
                     /gene="rep"
                     /product="replication enzyme"
                     /label="rep"
                     /note="derived from Clostridium perfringens plasmid pIP404
                     (INSD:M32882.1)"
                     /protein_id="BBD84638.1"
                     /translation="MRNALKLNKKNYIDNIHSRSKGWITRSVIDKKGYSQWHYKYAELK
                     DLDMSDENIYITLNTFYKPCRRLENIKELNTLFIDLDYYKTGKTKDQVLMDLEKNYFNQ
                     SIPIPNYVIDSGRGMYLIWIINAVPSKALPLWKAVQEYLYNQLKYFGADRQALDATRIL
                     RVPGSINSKSKTVVNILDEYEYIYDLREIQNGFLPELKPYERKKGRPSKINYIYRERSL
                     YYGRIQDIIKLCELREYDLKGHRELILFLYRYYLCSFTEDIEKALNDVLELNSMFRQHL
                     SEREVIRATRSAERCYLDKNKQYKYKNETLIELLEITEEEQKYMTIIISKKEYKRRENI
                     RGKKNYQEQLKAKGKATKKEELNVLRKKIKALKEKGFKNKEITLMLEVPIKTLERHITY
                     MKKNGLL"
     gene            complement(493..1713)
                     /gene="rep"
                     /label="rep"
     CDS             complement(2085..3245)
                     /codon_start=1
                     /gene="xylR"
                     /product="transcriptional repressor of the xylose operon"
                     /label="xylR"
                     /note="derived from Clostridium difficile ATCC 9689"
                     /protein_id="BBD84639.1"
                     /translation="MVTDKYTIREMNERLVLEQIIKNGPISRASIASTIGLNKATISAI
                     TKKLIDESLVHEIGIGNSTHSGGRKPILLVFNKCAGISLSMDVGYDYIFSSLSYLDGTI
                     INSKKLTDIQVSKDNVIQLIDEIINSYNISKIDTPYKVIGLTLAIHGITCENKVLFTPY
                     YNLNEIDLYSILSKKYDFPIHIENEANLTALAENTFSTVHNSLLSLSIHSGFGSGIIIN
                     NKLYSGRNGMSGEIGHTIIMPNGKLCPCGNRGCLEQYCSEKKVFEQLSSLENIPKIDSD
                     IVKQLYYEDNQNAKKVIHEFCSYLTIAINNAITTYAPEIIYLNSQIISDIPEILQITKD
                     MLVSSFNKGINIEISSLGSEASLYGGSAVNIKSFLNIQNLTLINEI"
     gene            complement(2085..3245)
                     /gene="xylR"
                     /label="xylR"
     CDS             3531..4244
                     /label="GFP (S65T)"
                     /note="S65T variant of Aequorea victoria green fluorescent
                     protein (Heim et al., 1995)"
     CDS             4245..4271
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             4272..4289
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(4605..5225)
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      5499..6087
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 aaaaaaattt ccaataatcc cactctaagc cacaaacacg ccctataaaa tcccgcttta
       61 atcccacttt gagacacatg taatattact ttacgcccta gtatagtgat aattttttac
      121 attcaatgcc acgcaaaaaa ataaaggggc actataataa aagttccttc ggaactaact
      181 aaagtaaaaa attatcttta caacctcccc aaaaaaaaga acaggtacaa agtaccctat
      241 aatacaagcg taaaaaaaat gagggtaaaa ataaaaaaat aaaaaaataa aaaaataaaa
      301 aaataaaaaa ataaaaaaat aaaaaaatat aaaaataaaa aaatataaaa ataaaaaaat
      361 ataaaaataa aaaaataaaa aaatataaaa ataaaaaaat aaaaaaatat aaaaatattt
      421 tttatttaaa gtttgaaaaa aattttttta tattatataa tctttgaaga aaagaatata
      481 aaaaatgagc ctttataaaa gcccattttt tttcatatac gtaatatgac gttctaatgt
      541 ttttattggt acttctaaca ttagagtaat ttctttattt ttaaagcctt tttctttaag
      601 ggcttttatt ttttttctta atacatttaa ttcctctttt tttgttgctt ttcctttagc
      661 ttttaattgc tcttgataat tttttttacc tctaatattt tctcttctct tatattcctt
      721 tttagaaatt attattgtca tatatttttg ttcttcttct gtaatttcta ataactctat
      781 aagagtttca ttcttatact tatattgctt atttttatct aaataacatc tttcagcact
      841 tctagttgct cttataactt ctctttcact taaatgttgt ctaaacatac tattaagttc
      901 taaaacatca tttaatgcct tctcaatgtc ttctgtaaag ctacaaagat aatatctata
      961 taaaaataat ataagctctc tgtgtccttt taaatcatat tctcttagtt cacaaagttt
     1021 tattatgtct tgtattcttc cataatataa acttctttct ctataaatat aatttatttt
     1081 gcttggtcta ccctttttcc tttcataagg ttttaattca ggtaaaaatc cattttgtat
     1141 ttctcttaag tcataaatat attcgtactc atctaatata ttgactactg tttttgattt
     1201 agagtttata cttcctggaa ctcttaatat tctggttgca tctaaggctt gtctatctgc
     1261 tccaaagtat tttaattgat tatataaata ttcttgaacc gctttccata atggtaatgc
     1321 tttactaggt actgcattta ttatccatat taaatacatt cctcttccac tatctattac
     1381 atagtttggt ataggaatac tttgattaaa ataattcttt tctaagtcca ttaatacctg
     1441 gtctttagtt ttgccagttt tataataatc caagtctata aacagtgtat ttaactcttt
     1501 tatattttct aatcgcctac acggcttata aaaggtattt agagttatat agatattttc
     1561 atcactcata tctaaatctt ttaattcagc gtatttatag tgccattggc tatatccttt
     1621 tttatctata acgctcctgg ttatccaccc tttacttcta ctatgaatat tatctatata
     1681 gttcttttta ttcagcttta atgcgtttct cacttattca cctccccttc tgtaaaacta
     1741 agaaaattat atcatatttt caataattat taactattct taaactctta ataaaaaata
     1801 gagtaagtcc ccaattgaaa cttaatctat tttttatgtt ttaatttatt atttttatta
     1861 aaatatttta aactaaatta aatgattctt tttaattttt tactatttca ttccataata
     1921 tattactata attatttaca aataatattt cttcatttgt aatatttaga tgatttacta
     1981 attttagtac tagtaggcct aggctaatct tgcataaact atataataaa tctaatttta
     2041 gaatgtataa tttaaagggt tatggctgta tatagtcata acccttatat ttcattaatt
     2101 aaagttaaat tttgtatatt caaaaagctc ttgatattta cggcagagcc accatataga
     2161 gatgcctcag accctaatga tgaaatttct atatttattc ctttgttaaa agagctaact
     2221 aacatatctt ttgtaatctg taatatctca gggatatcac taattatttg gctatttaaa
     2281 tatattattt caggagcata agttgtaata gcattattta tagctattgt taagtaacta
     2341 caaaactcat gaataacttt tttggcattc tggttatctt cataatatag ttgctttact
     2401 atatcagaat ctattttagg aatattttct aaagaagaaa gctgttcaaa tacctttttt
     2461 tctgaacaat actgttctaa acagccacga tttccacaag gacatagttt accattaggc
     2521 atgatgatag tgtgaccaat ttctccactc ataccatttc tgccactata taatttatta
     2581 tttattataa tgcctgaacc aaatccactg tgaatactga gactaagtag tgagttatgt
     2641 acagttgaaa aagtattctc agctaaagct gttaagtttg cttcattttc tatatgaata
     2701 ggaaagtcat actttttgct aaggatgcta tataagtcaa tctcattaag attgtaatat
     2761 ggagtaaaca atactttatt ttcacaagta atcccatgta tagccaaagt aagaccaatt
     2821 accttataag gagtgtctat tttggatatg ttataactat ttataatttc atctatcaat
     2881 tgtataacat tgtctttact tacttgtata tctgttaatt tcttagaatt tataatagtt
     2941 ccatctaagt aagatagaga agaaaatata tagtcgtatc caacgtccat acttaaagaa
     3001 atccctgcac atttattaaa tactaataat atgggttttc ttccgccact atgagtacta
     3061 tttccaattc ctatttcatg aactagagat tcatcaataa gttttttggt aatagcggat
     3121 atagttgctt tattcaatcc tatagtagaa gctatacttg ccctagaaat aggaccattt
     3181 tttataattt gttcaagtac caatctttca ttcatttctc gaatagtgta tttatcagta
     3241 accaatttga tattcctcct taaaataata ttgtaatact ttttacacaa aaataaaagg
     3301 ttattttgca ttgacaaaga taattaaata ttttattatt agttcataag ttagtttaat
     3361 atactaacaa aaataaagca agtaaaatat acctaaaata taaaaaaatt aggataggaa
     3421 aacgatagtt atgaagtggc attcaaggag ggatgcatat ggacgtcgac gcatgcatcg
     3481 atggatccgg taccgaattc ctgcagaagc ttgagctcga gtctagatct atgtcaaaag
     3541 gagaagaatt atttactgga gtagtaccaa tattagtaga attagatgga gatgtaaatg
     3601 gacataaatt ttcagtatca ggagaaggag aaggagatgc tacttatgga aaattaactt
     3661 taaaatttat atgtactact ggaaaattac cagtaccttg gccaacttta gtaactactt
     3721 taacttatgg agtacaatgt ttttcaagat atccagatca catgaaaaga catgattttt
     3781 ttaaatcagc tatgccagaa ggatatgtac aagaaagaac tatatttttt aaagatgatg
     3841 gaaattataa aactagagct gaagtaaaat ttgaaggaga tactttagta aatagaatag
     3901 aattaaaagg aatagatttt aaagaagatg gaaatatatt aggacataaa ttagaatata
     3961 attataattc acataatgta tatataatgg ctgataaaca aaaaaatgga ataaaagtaa
     4021 attttaaaat aagacataat atagaagatg gatcagtaca attagctgat cattatcaac
     4081 aaaatactcc aataggagat ggaccagtat tattaccaga taatcattat ttatcaactc
     4141 aatcagcttt atcaaaagat ccaaatgaaa aaagagatca catggtatta ttagaatttg
     4201 taactgctgc tggaataact catggaatgg atgaattata taaatatcct tatgatgttc
     4261 ctgattatgc tcatcatcat catcatcatt aattaattaa tattattaag atcatcttcg
     4321 gatgatcttt tttttgcagc tttaagctgc tttttttatt ttaaagtaaa ttttgaataa
     4381 atgtaggaga gtgttgggct agccccgggc ttggttgtgt tgcttttcgg ggcttaaatc
     4441 aaggaaaaat ccttgtttta agcctttcaa aaagaaacac aaggtctttg tactaacctg
     4501 tggttatgta taaaattgta gattttaggg taacaaaaaa caccgtattt ctacgatgtt
     4561 tttgcttaaa tacttgtttt tagttacaga caaacctgaa gttaactatt tatcaattcc
     4621 tgcaattcgt ttacaaaacg gcaaatgtga aatccgtcac atactgcgtg atgaacttga
     4681 attgccaaag gaagtataat tttgttatct tctttataat atttccccat agtaaaaata
     4741 ggaatcaaat aatcatatcc tttctgcaaa ttcagattaa agccatcgaa ggttgaccac
     4801 ggtatcatag atacattaaa aatgttttcc ggagcatttg gctttccttc cattctatga
     4861 ttgtttccat accgttgcgt atcactttca taatctgcta aaaatgattt aaagtcagac
     4921 ttacactcag tccaaaggct ggaaaatgtt tcagtatcat tgtgaaatat tgtatagctt
     4981 ggtatcatct catcatatat ccccaattca ccatcttgat tgattgccgt cctaaactct
     5041 gaatggcggt ttacaatcat tgcaatataa taaagcattg caggatatag tttcattccc
     5101 ttttccttta tttgtgtgat atccacttta acggtcatgc tgtatgtaca aggtacactt
     5161 gcaaagtagt ggtcaaaata ctcttttctg ttccaactat ttttatcaat tttttcaaat
     5221 accatctaag ttccctctca aattcaagtt tatcgctcta atgaacaaag atattatacc
     5281 acatttttgt gaatttttca acttgcccac ttcgactgca ctcccgactt aataacttct
     5341 tgaacacttg ccgaaaaaga aaaactgccg aaacttcatt tttaatttaa aaggatctag
     5401 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5641 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6121 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6181 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6241 agcgagtcag tgagcgagga agcggaagag cgcc
//
