LOCUS       V016462                 4856 bp    DNA     circular SYN 02-APR-2020
DEFINITION  Exported.
ACCESSION   V016462
VERSION     V016462
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4856)
  AUTHORS   Omura N, Yoshikawa T, Fujii H, Shibamura M, Inagaki T, Kato H, Egawa
            K, Harada S, Yamada S, Takeyama H, Saijo M.
  TITLE     A novel system for constructing a recombinant highly-attenuated
            vaccinia virus strain (LC16m8) expressing foreign genes and its
            application for the generation of LC16m8-based vaccines against
            herpes simplex virus 2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4856)
  AUTHORS   Yoshikawa T, Saijo M.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2017) Contact:Tomoki Yoshikawa National Institute
            of Infectious Diseases, Department of Virology I; 1-23-1 Toyama,
            Shinjuku-ku, Tokyo 162-8640, Japan URL
            :https://www.niid.go.jp/niid/en/
REFERENCE   3  (bases 1 to 4856)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-SEP-2017) Contact:Tomoki Yoshikawa National Institute of
            Infectious Diseases, Department of Virology I; 1-23-1 Toyama,
            Shinjuku-ku, Tokyo 162-8640, Japan URL
            :https://www.niid.go.jp/niid/en/"
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        239..257
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    281..724
                     /function="Homologous recombination site around B5R in VAC"
                     /label="B5R upstream"
                     /note="B5R upstream"
     CDS             complement(754..1209)
                     /gene="gpt"
                     /label="Xanthine-guanine phosphoribosyltransferase"
                     /note="Xanthine-guanine phosphoribosyltransferase from
                     Escherichia coli O17:K52:H18 (strain UMN026 / ExPEC).
                     Accession#: B7N8G9"
     regulatory      complement(1216..1255)
                     /label="VAC early late promoter"
                     /note="VAC early late promoter"
                     /regulatory_class="promoter"
     CDS             complement(1266..1973)
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     regulatory      complement(1974..2013)
                     /label="VAC early late promoter"
                     /note="VAC early late promoter"
                     /regulatory_class="promoter"
     misc_feature    2035..2279
                     /function="Intramolecular homologous recombination site to
                     B5R upstream"
                     /label="B5R upstream (short)"
                     /note="B5R upstream (short)"
     regulatory      2319..2358
                     /label="VAC early late promoter"
                     /note="VAC early late promoter"
                     /regulatory_class="promoter"
     misc_feature    2359..2364
                     /label="BglII site for insertion of gene of interest"
                     /note="BglII site for insertion of gene of interest"
     regulatory      2364..2370
                     /label="Vac early termination sequence"
                     /note="Vac early termination sequence"
                     /regulatory_class="terminator"
     promoter        complement(2402..2420)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2427..2443)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2543..3005
                     /function="Homologous recombination site around B5R in VAC"
                     /label="B5R downstream"
                     /note="B5R downstream"
     promoter        3042..3112
                     /label="AmpR promoter"
     CDS             3113..3970
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      4087..4675
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct ttgtacagtc gctacagcaa
      301 tataaatggt atattttaag atgtttcgat gaaatagata tcatgaagag atgttatata
      361 aaaaataaaa ctgtattcca attagttttt tgtatcaaag acattaatac tttaatgaga
      421 tacggtaaac atccttcttt cgtgaagtgc actagtctcg acgtatacgg aagtcgtgta
      481 cgtaatatca tagcatctat tagatatcgt cagagattaa ttagtctatt atccaagaag
      541 ctggatgcgg gagataaatg gtcgtgtttt cctaacgaaa taaaatataa aatattggaa
      601 aactttaacg ataacgaact atccacatat ctaaaaatct tataaacatt attaaaatat
      661 aaaatctaag tggataaaat cacactacat cattgtttcc ttttagtgct cgacagtgta
      721 agcttggtac cgagctcgga tccataaaaa ttagcgaccg gagattggcg ggacgaatac
      781 gacgcccata tcccacggct gttcaatcca ggtatcttgc gggatatcaa caacatagtc
      841 atcaaccagc ggacgaccag ccggttttgc gaagatggtg acaaagtgcg cttttggata
      901 catttcacga atcgcaaccg cagtaccacc ggtatccacc aggtcatcaa taacgatgaa
      961 gccttcgcca tcgccttctg cgcgtttcag cactttaagc tcgcgctggt tgtcgtgatc
     1021 gtagctggaa atacaaacgg tatcgacatg acgaataccc agttcacgcg ccagtaacgc
     1081 acccggtacc agaccgccac ggcttacggc aataatgcct ttccattgtt cagaaggcat
     1141 cagtcggctt gcgagtttac gtgcatggat ctgcaacatg tcccaggtga cgatgtattt
     1201 ttcgctcatg gtggctattt atattccaaa aaaaaaaaat aaaatttcaa tttttataaa
     1261 aattacttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgctcgtac
     1321 tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccagcttgat gttgacgttg
     1381 taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcggcgtcg
     1441 tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc
     1501 tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg
     1561 ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc
     1621 agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc
     1681 cacttgaagc cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc
     1741 acgtaggcct tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg
     1801 gggccgccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg
     1861 ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg caccttgaag
     1921 cgcatgaact ccttgatgat ggccatgttg tcctcctcgc ccttgctcac cattatttat
     1981 attccaaaaa aaaaaaataa aatttcaatt tttaccggtc ccaagggcga attctgcagc
     2041 gtaatatcat agcatctatt agatatcgtc agagattaat tagtctatta tccaagaagc
     2101 tggatgcggg agataaatgg tcgtgttttc ctaacgaaat aaaatataaa atattggaaa
     2161 actttaacga taacgaacta tccacatatc taaaaatctt ataaacatta ttaaaatata
     2221 aaatctaagt ggataaaatc acactacatc attgtttcct tttagtgctc gacagtgtct
     2281 gcagatatcc atcacactgg cggccgctcg aagtcgacaa aaattgaaat tttatttttt
     2341 ttttttggaa tataaataag atctttttat ctcgagcatg catctagagg gcccaattcg
     2401 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2461 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2521 aatagcgaag aggcccgcac cgatcgagtg gcgttaacaa ttatgggcgt catattttta
     2581 atctccgtta tagtattagt ttgttcctgt gacaaaaata atgaccaata taagttccat
     2641 aaattgctac cgtaaatata aatccgttaa aataattaat aattaataac gaacaagtat
     2701 caaaagatta aagacttata gctagaatca attgagatgt cttcttcagt ggatgttgat
     2761 atctacgatg ccgttagagc atttttactc aggcactatt ataacaagag atttattgtg
     2821 tatggaagaa gtaacgccat attacataat atatacaggc tatttacaag atgcgccgtt
     2881 ataccgttcg atgatatagt acgtactatg ccaaatgaat cacgtgttaa acaatgggtg
     2941 atggatacac ttaatggtat aatgatgaat gaacgcgatg tttctgtaag cgttggcacc
     3001 ggacgatcgc ccttcccaac agttgcgcag cctatacgta cgttcaaata tgtatccgct
     3061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3241 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3301 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3361 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3421 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3481 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3541 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3601 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3661 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3721 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3781 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3841 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3901 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3961 taagcattgg taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4681 cagcaacgcg gcctttttac ggttcctggg cttttgctgg ccttttgctc acatgttctt
     4741 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4801 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
