From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Plasmid pH2515, shuttle vector for E.coli and L.casei. complete
Date: 21 Jul 92 10:08:42 GMT
Distribution: bionet
Checksum: 24102    17

LOCUS       SYNPH2515    6550 bp ds-DNA   Circular  SYN       21-JUL-1992
DEFINITION  Plasmid pH2515, shuttle vector for E.coli and L.casei. complete
            sequence.
ACCESSION   D90381
KEYWORDS    ampicillin resistance;
            macrolide-lincosamide-streptogramin B resistance; shuttle vector;
            tetracycline resistance.
SOURCE      Plasmid pH2515 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 51 to 1160)
  AUTHORS   Brehm,J., Salmond,G. and Minton,N.
  TITLE     Sequence of the adenine methylase gene of the Streptococcus
            faecalis plasmid pAMbeta1
  JOURNAL   Nucleic Acids Res. 15, 3177-3177 (1987)
  STANDARD  simple automatic
REFERENCE   3  (bases 1 to 43; 1709 to 6550)
  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  JOURNAL   Unpublished (1991)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
REFERENCE   4  (bases 1 to 6550)
  AUTHORS   Shimizu-Kadota,M., Shibahara-Sone,H. and Ishiwa,H.
  TITLE     Shuttle plasmid vectors for Lactobacillus casei and Escherichia
            coli with a minus origin
  JOURNAL   Unpublished (1991)
  STANDARD  full staff_review
COMMENT
            [1]  sites.
FEATURES             Location/Qualifiers
     misc_feature    1..42
                     /note="poly linker"
     misc_feature    1..42
                     /note="poly linker"
     -35_signal      223..228
     -35_signal      223..228
     -10_signal      250..255
     -10_signal      250..255
     RBS             298..303
     misc_binding    298..303
                     /note="ribosome binding site"
     CDS             311..1048
                     /note="MLS resistance protein"
                     /codon_start=1
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLT
                     TKLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIP
                     YHLSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAE
                     CFHPKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHA
                     KVNNLSTVTYEQVLSIFNSYLLFNGRK"
     CDS             311..1048
                     /product="MLS resistance protein"
                     /codon_start=1
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLT
                     TKLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIP
                     YHLSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAE
                     CFHPKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHA
                     KVNNLSTVTYEQVLSIFNSYLLFNGRK"
     rep_origin      1451..1505
                     /note="minus origin for plasmid replication replication in
                     Gram-positive bacteria"
     rep_origin      1451..1505
                     /note="minus origin for plasmid replication replication in
                     Gram-positive bacteria"
     CDS             complement(1779..3158)
                     /note="Tc resistance protein, gtg start codon"
                     /codon_start=1
                     /translation="NVNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIAND
                     FNKPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHS
                     FFSLLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIG
                     GMIAHYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLF
                     TTSYSISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGF
                     VSMVPYMMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFL
                     SVSFLTASFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNF
                     TSFLSEGTGIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLN
                     VYKHSQRDF"
                     /partial
     CDS             complement(1779..3155)
                     /note="gtg start codon"
                     /product="Tc resistance protein"
                     /codon_start=1
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDF
                     NKPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSF
                     FSLLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGG
                     MIAHYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFT
                     TSYSISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFV
                     SMVPYMMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLS
                     VSFLTASFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFT
                     SFLSEGTGIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNV
                     YKHSQRDF"
     RBS             complement(3163..3167)
     misc_binding    complement(3163..3167)
                     /note="ribosome binding site"
     -10_signal      complement(3288..3293)
     -10_signal      complement(3288..3293)
     -35_signal      complement(3311..3316)
     -35_signal      complement(3311..3316)
     CDS             complement(3362..4348)
                     /note="Rep-alpha-1 (replication protein)"
                     /codon_start=1
                     /translation="IMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLH
                     ILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQK
                     VVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFM
                     RATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVK
                     VQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISY
                     GGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
                     /partial
     CDS             complement(3362..4345)
                     /product="Rep-alpha-1 (replication protein)"
                     /codon_start=1
                     /translation="MCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDYFELLHI
                     LEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGIQSQKV
                     VAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLVGFMR
                     ATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPNVKV
                     QMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLISYG
                     GLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     RBS             complement(4360..4364)
     misc_binding    complement(4360..4364)
                     /note="ribosome binding site"
     -10_signal      complement(4448..4454)
     -10_signal      complement(4448..4454)
     -35_signal      complement(4471..4476)
     -35_signal      complement(4471..4476)
     -10_signal      complement(4499..4505)
     -10_signal      complement(4499..4505)
     -35_signal      complement(4520..4527)
     -35_signal      complement(4520..4527)
     rep_origin      complement(4524..4525)
                     /note="replication origin in Gram-positive bacteria (nick
                     site)"
     rep_origin      complement(4524..4525)
                     /note="replication origin in Gram-positive bacteria (nick
                     site)"
     -35_signal      4898..4903
     -35_signal      4898..4903
     -10_signal      4921..4926
     -10_signal      4921..4926
     RBS             4957..4964
     misc_binding    4957..4964
                     /note="ribosome binding site"
     CDS             4968..5828
                     /note="Ap resistance protein"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     CDS             4968..5828
                     /product="Ap resistance protein"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     misc_feature    6029..6539
                     /note="RNA primer"
     misc_feature    6029..6539
                     /note="RNA primer"
BASE COUNT     2069 a   1332 c   1216 g   1933 t
ORIGIN
        1 aagcttctag agatctgcag gtcgacggat ccccgggaat tccccgcaca aaaagaaaaa
       61 cgaaatgata caccaatcag tgcaaaaaaa gatataatgg gagataagac ggttcgtgtt
      121 cgtgctgact tgcaccatat cataaaaatc gaaacagcaa agaatggcgg aaacgtaaaa
      181 gaagttatgg aaataagact tagaagcaaa cttaagagtg tgttgatagt gcagtatctt
      241 aaaattttgt ataataggaa ttgaagttaa attagatgct aaaaatttgt aattaagaag
      301 gagtgattac atgaacaaaa atataaaata ttctcaaaac tttttaacga gtgaaaaagt
      361 actcaaccaa ataataaaac aattgaattt aaaagaaacc gataccgttt acgaaattgg
      421 aacaggtaaa gggcatttaa cgacgaaact ggctaaaata agtaaacagg taacgtctat
      481 tgaattagac agtcatctat tcaacttatc gtcagaaaaa ttaaaactga atactcgtgt
      541 cactttaatt caccaagata ttctacagtt tcaattccct aacaaacaga ggtataaaat
      601 tgttgggagt attccttacc atttaagcac acaaattatt aaaaaagtgg tttttgaaag
      661 ccatgcgtct gacatctatc tgattgttga agaaggattc tacaagcgta ccttggatat
      721 tcaccgaaca ctagggttgc tcttgcacac tcaagtctcg attcagcaat tgcttaagct
      781 gccagcggaa tgctttcatc ctaaaccaaa agtaaacagt gtcttaataa aacttacccg
      841 ccataccaca gatgttccag ataaatattg gaagctatat acgtactttg tttcaaaatg
      901 ggtcaatcga gaatatcgtc aactgtttac taaaaatcag tttcatcaag caatgaaaca
      961 cgccaaagta aacaatttaa gtaccgttac ttatgagcaa gtattgtcta tttttaatag
     1021 ttatctatta tttaacggga ggaaataatt ctatgagtcg cttttgtaaa tttggaaagt
     1081 tacacgttac taaagggaat gtagataaat tattaggtat actactgaca gcttccaagg
     1141 agctaaagag gtccctagcg ggggatctag gcgccttgca ccataaaatt gtaaacgtta
     1201 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1261 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg
     1321 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1381 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg
     1441 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1501 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1561 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1621 atgcgccgct acagggcgcg tactatggtt gctttgacga gcgcggtgtg aaataccgca
     1681 cagatgcgta aggagaaaat accgcatcag gcgcctaatt cctgttataa aaaaaggatc
     1741 aattttgaac tctctcccaa agttgatccc ttaacgattt agaaatccct ttgagaatgt
     1801 ttatatacat tcaaggtaac cagccaacta atgacaatga ttcctgaaaa aagtaataac
     1861 aaattactat acagataagt tgactgatca acttccatag gtaacaacct ttgatcaagt
     1921 aagggtatgg ataataaacc acctacaatt gcaatacctg ttccctctga taaaaagctg
     1981 gtaaagttaa gcaaactcat tccagcacca gcttcctgct gtttcaagct acttgaaaca
     2041 attgttgata taactgtttt ggtgaacgaa agcccaccta aaacaaatac gattataatt
     2101 gtcatgaacc atgatgttgt ttctaaaaga aaggaagcag ttaaaaagct aacagaaaga
     2161 aatgtaactc cgatgtttaa cacgtataaa ggacctcttc tatcaacaag tatcccacca
     2221 atgtagccga aaataatgac actcattgtt ccagggaaaa taattacact tccgatttcg
     2281 gcagtactta gctggtgaac atctttcatc atataaggaa ccatagagac aaaccctgct
     2341 actgttccaa atataattcc cccacaaaga actccaatca taaaaggtat atttttccct
     2401 aatccgggat caacaaaagg atctgttact ttcctgatat gttttacaaa tatcaggaat
     2461 gacagcacgc taacgataag aaaagaaatg ctatatgatg ttgtaaacaa cataaaaaat
     2521 acaatgccta cagacattag tataattcct ttgatatcaa aatgaccttt tatccttact
     2581 tctttcttta ataatttcat aagaaacgga acagtgataa ttgttatcat aggaatgagt
     2641 agaagatagg accaatgaat ataatgggct atcattccac caatcgctgg accgactcct
     2701 tctcccatgg ctactatcga tccaataaga ccaaatgctt tacccctatt ttcctttgga
     2761 atatagcgcg caactacaac cattacgagt gctggaaatg cagctgcacc agccccttga
     2821 ataaaacgag ccataataag taaggaaaag aaagaatggc caacaaaccc aattaccgac
     2881 ccgaaacaat ttattataat tccaaatagg agtaaccttt tgatgcctaa ttgatcagat
     2941 agctttccat atacagctgt tccaatggaa aaggttaaca taaaggctgt gttcacccag
     3001 tttgtactcg caggtggttt attaaaatca tttgcaatat caggtaatga gacgttcaaa
     3061 accatttcat ttaatacgct aaaaaaagat aaaatgcaaa gccaaattaa aatttggttg
     3121 tgtcgtaaat tcgattgtga ataggatgta ttcacatttc accctccaat aatgagggca
     3181 gacgtagttt atagggttaa tgatacgctt ccctctttta attgaaccct gttacattca
     3241 ttattcatta cacttcataa ttaattcctc ctaaacttga ttaaaacatt ttaccacata
     3301 taaactaagt tttaaattca gtatttcatc acttatacaa caatatggcc cgtttgttga
     3361 actactcttt aataaaataa tttttccgtt cccaattcca cattgcaata atagaaaatc
     3421 catcttcatc ggctttttcg tcatcatctg tatgaatcaa atcgccttct tctgtgtcat
     3481 caaggtttaa ttttttatgt atttctttta acaaaccacc ataggagatt aaccttttac
     3541 ggtgtaaacc ttcctccaaa tcagacaaac gtttcaaatt cttttcttca tcatcggtca
     3601 taaaatccgt atcctttaca ggatattttg cagtttcgtc aattgccgat tgtatatccg
     3661 atttatattt atttttcggt cgaatcattt gaacttttac atttggatca tagtctaatt
     3721 tcattgcctt tttccaaaat tgaatccatt gtttttgatt cacgtagttt tctgtattct
     3781 taaaataagt tggttccaca cataccaata catgcatgtg ctgattataa gaattatctt
     3841 tattatttat tgtcacttcc gttgcacgca taaaaccaac aagattttta ttaatttttt
     3901 tatattgcat cattcggcga aatccttgag ccatatctga caaactctta tttaattctt
     3961 cgccatcata aacattttta actgttaatg tgagaaacaa ccaacgaact gttggctttt
     4021 gtttaataac ttcagcaaca accttttgtg actgaatgcc atgtttcatt gctctcctcc
     4081 agttgcacat tggacaaagc ctggatttac aaaaccacac tcgatacaac tttctttcgc
     4141 ctgtttcacg attttgttta tactctaata tttcagcaca atcttttact ctttcagcct
     4201 ttttaaattc aagaatatgc agaagttcaa agtaatcaac attagcgatt ttcttttctc
     4261 tccatggtct cacttttcca ctttttgtct tgtccactaa aacccttgat ttttcatctg
     4321 aataaatgct actattagga cacataatat taaaagaaac ccccatctat ttagttattt
     4381 gtttggtcac ttataacttt aacagatggg gtttttctgt gcaaccaatt ttaagggttt
     4441 tcaatacttt aaaacacata cataccaaca cttcaacgca cctttcagca actaaaataa
     4501 aaatgacgtt atttctatat gtatcaagat aagaaagaac aagttcaaaa ccatcaaaaa
     4561 aagacacctt ttcaggtgct ttttttattt tataaactca ttccctgatc tcgacttcgt
     4621 tcttttttta cctctcggtt atgagttagt tcaaattcgt tctttttagg ttctaaatcg
     4681 tgtttttctt ggaattgtgc tgttttatcc tttaccttgt ctacaaaccc cttaaaaacg
     4741 tttttaaagg cttttaagcc gtctgtacgt tccttaagga attattcctt agtgctttct
     4801 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4861 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4921 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4981 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5041 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5101 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5161 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     5221 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5281 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5341 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5401 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5461 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gccgcaatgg
     5521 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5581 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5641 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5701 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5761 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5821 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5881 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5941 aacgtgagtt ttcgttccac tgagcgtcag accccttaat aagatgatct tcttgagatc
     6001 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggaggtt
     6061 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
     6121 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
     6181 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
     6241 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
     6301 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     6361 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     6421 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6481 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg ggcggagcct atggaaaaac
     6541 ggctttgccc
//