From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Plasmid pHY320PLK, shuttle vector for E.coli and B.subtilis,
Date: 21 Jul 92 10:08:40 GMT
Distribution: bionet
Checksum: 52256    12

LOCUS       SYN320PLK    5387 bp ds-DNA   Circular  SYN       21-JUL-1992
DEFINITION  Plasmid pHY320PLK, shuttle vector for E.coli and B.subtilis,
            complete sequence.
ACCESSION   D00946 D00054
KEYWORDS    amipicillin resistance; shuttle vector; tetracycline resistance.
SOURCE      Plasmid pHY320PLK DNA from E.faecalis (strain DS-5) and E.coli.
  ORGANISM  Artificial gene
            Artificial sequences; Genes.
REFERENCE   1  (sites)
  AUTHORS   Ishiwa,H. and Shibahara,H.
  TITLE     New shuttle vectors for Escherichia coli and Bacillus subtilis.
            III. Nucleotide sequence analysis of tetracycline resistance gene
            of pAMalpha1 and ori-177
  JOURNAL   Jpn. J. Genet. 60, 485-498 (1985)
  STANDARD  full staff_review
REFERENCE   2  (bases 1 to 43; 554 to 5386)
  AUTHORS   Ishiwa,H. and Shibahara-Sone,H.
  TITLE     New shuttle vectors for Escherichia coli and Bacillus subtilis. IV.
            The nucleotide sequence of pHY300PLK and some properties in
            relation to transformation
  JOURNAL   Jpn. J. Genet. 61, 515-528 (1986)
  STANDARD  full staff_entry
REFERENCE   3  (bases 1 to 5387)
  AUTHORS   Ishiwa,H., Shibahara-Sone,H. and Shimizu-Kadota,M.
  TITLE     New shuttle vectors for Escherichia coli and Bacillus subtilis. V
  JOURNAL   Unpublished (1991)
  STANDARD  full staff_review
COMMENT
            [1]  sites.
FEATURES             Location/Qualifiers
     misc_feature    1..51
                     /note="polylinker"
     rep_origin      102..519
                     /note="BA3 minus origin"
     CDS             complement(616..1992)
                     /note="gtg start codon"
                     /product="tetracycline resistance protein"
                     /codon_start=1
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDF
                     NKPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSF
                     FSLLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGG
                     MIAHYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFT
                     TSYSISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFV
                     SMVPYMMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLS
                     VSFLTASFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFT
                     SFLSEGTGIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNV
                     YKHSQRDF"
     RBS             complement(2000..2004)
     -10_signal      complement(2125..2130)
     -35_signal      complement(2148..2153)
     CDS             complement(2199..3182)
                     /product="Rep-alpha-1 (replication protein)"
                     /codon_start=1
                     /translation="MCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLHI
                     LEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQKV
                     VAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFMR
                     ATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVKV
                     QMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISYG
                     GLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     RBS             complement(3197..3201)
     variation       3278..3279
                     /note="tc in [3]; tcc in [2]"
     -10_signal      complement(3285..3291)
     -35_signal      complement(3308..3313)
     -10_signal      complement(3336..3342)
     -35_signal      complement(3357..3364)
     rep_origin      complement(3361..3362)
                     /note="replication origin in Gram-positive bacteria (nick
                     site)"
     variation       3596
                     /note="c in [3]; deletion in [2]"
     variation       3699
                     /note="g in [3]; c in [2]"
     -35_signal      3735..3740
     -10_signal      3758..3763
     RBS             3794..3801
     CDS             3805..4665
                     /product="ampicillin resistance protein"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     variation       4350
                     /note="c in [3]; a in [2]"
     misc_feature    4866..5376
                     /note="RNA primer"
     variation       4949
                     /note="g in [3]; c in [2]"
     variation       5378
                     /note="g in [3]; deletion in [2]"
BASE COUNT     1625 a   1138 c    961 g   1663 t
ORIGIN
        1 aagcttctag agatctgcag gtcgacggat ccccgggaat tccggagcgc tcctccccct
       61 tttttttctt gtgtcgttcg gtgtctttgg atttcggcag tacttccaag taaagtataa
      121 cacactatac tttatattca taaagtgtgt gctctgcgag gctgtcggca gtgccgacca
      181 aaaccataaa acctttaaga cctttctttt ttttacgaga aaaaagaaac aaaaaaacct
      241 gccctctgcc acctcagcaa aggggggttt tgctctcgtg ctcgtttaaa aatcagcaag
      301 ggacaggtag tattttttga gaagatcact caaaaaatct ccacctttaa acccttgcca
      361 atttttattt tgtccgtttt gtctagctta ccgaaagcca gactcagcaa gaataaaatt
      421 tttattgtct ttcggttttc tagtgtaacg gacaaaacca ctcaaaataa aaaagataca
      481 agagaggtct ctcgtatctt ttattcagca atcgcgccct ttaatggaac aaaaagattt
      541 gaattaggcg cctaattcct gttataaaaa aaggatcaat tttgaactct ctcccaaagt
      601 tgatccctta acgatttaga aatccctttg agaatgttta tatacattca aggtaaccag
      661 ccaactaatg acaatgattc ctgaaaaaag taataacaaa ttactataca gataagttga
      721 ctgatcaact tccataggta acaacctttg atcaagtaag ggtatggata ataaaccacc
      781 tacaattgca atacctgttc cctctgataa aaagctggta aagttaagca aactcattcc
      841 agcaccagct tcctgctgtt tcaagctact tgaaacaatt gttgatataa ctgttttggt
      901 gaacgaaagc ccacctaaaa caaatacgat tataattgtc atgaaccatg atgttgtttc
      961 taaaagaaag gaagcagtta aaaagctaac agaaagaaat gtaactccga tgtttaacac
     1021 gtataaagga cctcttctat caacaagtat cccaccaatg tagccgaaaa taatgacact
     1081 cattgttcca gggaaaataa ttacacttcc gatttcggca gtacttagct ggtgaacatc
     1141 tttcatcata taaggaacca tagagacaaa ccctgctact gttccaaata taattccccc
     1201 acaaagaact ccaatcataa aaggtatatt tttccctaat ccgggatcaa caaaaggatc
     1261 tgttactttc ctgatatgtt ttacaaatat caggaatgac agcacgctaa cgataagaaa
     1321 agaaatgcta tatgatgttg taaacaacat aaaaaataca atgcctacag acattagtat
     1381 aattcctttg atatcaaaat gaccttttat ccttacttct ttctttaata atttcataag
     1441 aaacggaaca gtgataattg ttatcatagg aatgagtaga agataggacc aatgaatata
     1501 atgggctatc attccaccaa tcgctggacc gactccttct cccatggcta ctatcgatcc
     1561 aataagacca aatgctttac ccctattttc ctttggaata tagcgcgcaa ctacaaccat
     1621 tacgagtgct ggaaatgcag ctgcaccagc cccttgaata aaacgagcca taataagtaa
     1681 ggaaaagaaa gaatggccaa caaacccaat taccgacccg aaacaattta ttataattcc
     1741 aaataggagt aaccttttga tgcctaattg atcagatagc tttccatata cagctgttcc
     1801 aatggaaaag gttaacataa aggctgtgtt cacccagttt gtactcgcag gtggtttatt
     1861 aaaatcattt gcaatatcag gtaatgagac gttcaaaacc atttcattta atacgctaaa
     1921 aaaagataaa atgcaaagcc aaattaaaat ttggttgtgt cgtaaattcg attgtgaata
     1981 ggatgtattc acatttcacc ctccaataat gagggcagac gtagtttata gggttaatga
     2041 tacgcttccc tcttttaatt gaaccctgtt acattcatta ttcattacac ttcataatta
     2101 attcctccta aacttgatta aaacatttta ccacatataa actaagtttt aaattcagta
     2161 tttcatcact tatacaacaa tatggcccgt ttgttgaact actctttaat aaaataattt
     2221 ttccgttccc aattccacat tgcaataata gaaaatccat cttcatcggc tttttcgtca
     2281 tcatctgtat gaatcaaatc gccttcttct gtgtcatcaa ggtttaattt tttatgtatt
     2341 tcttttaaca aaccaccata ggagattaac cttttacggt gtaaaccttc ctccaaatca
     2401 gacaaacgtt tcaaattctt ttcttcatca tcggtcataa aatccgtatc ctttacagga
     2461 tattttgcag tttcgtcaat tgccgattgt atatccgatt tatatttatt tttcggtcga
     2521 atcatttgaa cttttacatt tggatcatag tctaatttca ttgccttttt ccaaaattga
     2581 atccattgtt tttgattcac gtagttttct gtattcttaa aataagttgg ttccacacat
     2641 accaatacat gcatgtgctg attataagaa ttatctttat tatttattgt cacttccgtt
     2701 gcacgcataa aaccaacaag atttttatta atttttttat attgcatcat tcggcgaaat
     2761 ccttgagcca tatctgacaa actcttattt aattcttcgc catcataaac atttttaact
     2821 gttaatgtga gaaacaacca acgaactgtt ggcttttgtt taataacttc agcaacaacc
     2881 ttttgtgact gaatgccatg tttcattgct ctcctccagt tgcacattgg acaaagcctg
     2941 gatttacaaa accacactcg atacaacttt ctttcgcctg tttcacgatt ttgtttatac
     3001 tctaatattt cagcacaatc ttttactctt tcagcctttt taaattcaag aatatgcaga
     3061 agttcaaagt aatcaacatt agcgattttc ttttctctcc atggtctcac ttttccactt
     3121 tttgtcttgt ccactaaaac ccttgatttt tcatctgaat aaatgctact attaggacac
     3181 ataatattaa aagaaacccc catctattta gttatttgtt tggtcactta taactttaac
     3241 agatggggtt tttctgtgca accaatttta agggttttca atactttaaa acacatacat
     3301 accaacactt caacgcacct ttcagcaact aaaataaaaa tgacgttatt tctatatgta
     3361 tcaagataag aaagaacaag ttcaaaacca tcaaaaaaag acaccttttc aggtgctttt
     3421 tttattttat aaactcattc cctgatctcg acttcgttct ttttttacct ctcggttatg
     3481 agttagttca aattcgttct ttttaggttc taaatcgtgt ttttcttgga attgtgctgt
     3541 tttatccttt accttgtcta caaacccctt aaaaacgttt ttaaaggctt ttaagccgtc
     3601 tgtacgttcc ttaaggaatt attccttagt gctttctagg ttaatgtcat gataataatg
     3661 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3781 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3841 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3901 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3961 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4021 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     4081 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4141 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4201 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4261 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4321 aacgacgagc gtgacaccac gatgcctgcc gcaatggcaa caacgttgcg caaactatta
     4381 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4441 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4501 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4561 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4621 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4801 gcgtcagacc ccttaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     4861 tgctctgaaa acgaaaaaac cgccttgcag ggaggttttt cgaaggttct ctgagctacc
     4921 aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     4981 gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     5041 tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     5101 taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     5161 tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     5221 tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     5281 acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     5341 cgtgatgctt gtcagggggc ggagcctatg gaaaaacggc tttgccc
//