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ID   PHY320PLK  preliminary; circular DNA; SYN; 5387 BP.
XX
AC   D00946; D00054; N00054;
XX
DT   19-SEP-1987 (Rel. 2, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   Bacillus/E.coli plasmid vector pHY320PLK - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-43; 554-5386
RC   pHY300.1PLK, pHY300.2PLK from pHY300PLK & linker
RC   pHY300.8PLK, pHY300.9PLK from pHY300PLK & lambda
RA   Ishiwa H., Shibahara-Sone H.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis.
RT   IV. The nucleotide sequence of pHY300PLK and some properties in
RT   relation to transformation";
RL   Jpn. J. Genet. 61:515-528(1986).
XX
RN   [2]
RP   1-5387
RC   pHY320PLK from pHY300PLK
RA   Ishiwa H., Shibahara-Sone H., Shimizu-Kadota M.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis. V.";
RL   Unpublished (1992).
XX
CC   PHY320PLK is one of the smallest hybrids of a new series of
CC   chimeric plasmids using the parental plasmids, pACYC177 of E. coli
CC   and pAM-alpha of Streptococcus faecalis.
CC   This shuttle vector for E. coli and B. subtilis contains an
CC   RNA primer gene for ori-177; the R-Tc gene; the R-Ap gene;
CC   the Rep-alpha-1 gene, which is the plasmid replication gene;
CC   two replication origins, one for E.coli and the other for B. subtilis;
CC   and a polylinker.
CC   Twenty-three unique recognition restriction endonuclease sites have
CC   been found in pHY320PLK.
CC   NM (pHY320PLK)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)(B.subtilis)(E.faecalis strain DS-5)(S.faecalis)
CC   CP ()
CC   FN (shuttle)(transformation)
CC   SE ()
CC   PA (pACYC177)(pAMalpha)(pHY300PLK)
CC   BR (pHY163PLK)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pHY300PLK 4870bp
FT                   -> pHY320PLK 5387bp"
FT   misc_binding    1..51
FT                   /note="MCS"
FT   rep_origin      102..519
FT                   /note="ORI Streptococcus faecalis BA3 minus origin"
FT   CDS             complement(616..1992)
FT                   /note="ANT Gram-positive bacteria tetracycline
FT                   resistance gene; gtg start codon"
FT   RBS             complement(2000..2004)
FT                   /note="RBS Gram-positive bacteria tetracycline
FT                   resistance gene"
FT   -10_signal      complement(2125..2130)
FT                   /note="S10 Gram-positive bacteria tetracycline
FT                   resistance gene"
FT   -35_signal      complement(2148..2153)
FT                   /note="S35 Gram-positive bacteria tetracycline
FT                   resistance gene"
FT   CDS             complement(2199..3182)
FT                   /note="GEN Gram-positive bacteria Rep-alpha-1 gene
FT                   (replication protein)"
FT   RBS             complement(3197..3201)
FT                   /note="RBS Gram-positive bacteria Rep-alpha-1 gene"
FT   variation       3278..3279
FT                   /note="tc in [3]; tcc in [2]"
FT   -10_signal      complement(3285..3291)
FT                   /note="S10 Gram-positive bacteria Rep-alpha-1 gene"
FT   -35_signal      complement(3308..3313)
FT                   /note="S35 Gram-positive bacteria Rep-alpha-1 gene"
FT   -10_signal      complement(3336..3342)
FT                   /note="S10 Gram-positive bacteria Rep-alpha-1 gene"
FT   -35_signal      complement(3357..3364)
FT                   /note="S35 Gram-positive bacteria Rep-alpha-1 gene"
FT   rep_origin      complement(3361..3362)
FT                   /note="ORI Gram-positive bacteria replication origin
FT                   (nick site)"
FT   variation       3596..3596
FT                   /note="c in [3]; deletion in [2]"
FT   variation       3699..3699
FT                   /note="g in [3]; c in [2]"
FT   -35_signal      3735..3740
FT                   /note="S35 E. coli beta-lactamase gene"
FT   -10_signal      3758..3763
FT                   /note="S10 E. coli beta-lactamase gene"
FT   RBS             3794..3801
FT                   /note="RBS E. coli beta-lactamase gene"
FT   CDS             3805..4665
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   variation       4350..4350
FT                   /note="c in [3]; a in [2]"
FT   misc_feature    4866..5376
FT                   /note="RNA primer, from ori-177"
FT   variation       4949..4949
FT                   /note="g in [3]; c in [2]"
FT   variation       5378..5378
FT                   /note="g in [3]; deletion in [2]"
XX
SQ   Sequence 5387 BP; 1625 A; 1138 C; 961 G; 1663 T; 0 other;
     aagcttctag agatctgcag gtcgacggat ccccgggaat tccggagcgc tcctccccct
     tttttttctt gtgtcgttcg gtgtctttgg atttcggcag tacttccaag taaagtataa
     cacactatac tttatattca taaagtgtgt gctctgcgag gctgtcggca gtgccgacca
     aaaccataaa acctttaaga cctttctttt ttttacgaga aaaaagaaac aaaaaaacct
     gccctctgcc acctcagcaa aggggggttt tgctctcgtg ctcgtttaaa aatcagcaag
     ggacaggtag tattttttga gaagatcact caaaaaatct ccacctttaa acccttgcca
     atttttattt tgtccgtttt gtctagctta ccgaaagcca gactcagcaa gaataaaatt
     tttattgtct ttcggttttc tagtgtaacg gacaaaacca ctcaaaataa aaaagataca
     agagaggtct ctcgtatctt ttattcagca atcgcgccct ttaatggaac aaaaagattt
     gaattaggcg cctaattcct gttataaaaa aaggatcaat tttgaactct ctcccaaagt
     tgatccctta acgatttaga aatccctttg agaatgttta tatacattca aggtaaccag
     ccaactaatg acaatgattc ctgaaaaaag taataacaaa ttactataca gataagttga
     ctgatcaact tccataggta acaacctttg atcaagtaag ggtatggata ataaaccacc
     tacaattgca atacctgttc cctctgataa aaagctggta aagttaagca aactcattcc
     agcaccagct tcctgctgtt tcaagctact tgaaacaatt gttgatataa ctgttttggt
     gaacgaaagc ccacctaaaa caaatacgat tataattgtc atgaaccatg atgttgtttc
     taaaagaaag gaagcagtta aaaagctaac agaaagaaat gtaactccga tgtttaacac
     gtataaagga cctcttctat caacaagtat cccaccaatg tagccgaaaa taatgacact
     cattgttcca gggaaaataa ttacacttcc gatttcggca gtacttagct ggtgaacatc
     tttcatcata taaggaacca tagagacaaa ccctgctact gttccaaata taattccccc
     acaaagaact ccaatcataa aaggtatatt tttccctaat ccgggatcaa caaaaggatc
     tgttactttc ctgatatgtt ttacaaatat caggaatgac agcacgctaa cgataagaaa
     agaaatgcta tatgatgttg taaacaacat aaaaaataca atgcctacag acattagtat
     aattcctttg atatcaaaat gaccttttat ccttacttct ttctttaata atttcataag
     aaacggaaca gtgataattg ttatcatagg aatgagtaga agataggacc aatgaatata
     atgggctatc attccaccaa tcgctggacc gactccttct cccatggcta ctatcgatcc
     aataagacca aatgctttac ccctattttc ctttggaata tagcgcgcaa ctacaaccat
     tacgagtgct ggaaatgcag ctgcaccagc cccttgaata aaacgagcca taataagtaa
     ggaaaagaaa gaatggccaa caaacccaat taccgacccg aaacaattta ttataattcc
     aaataggagt aaccttttga tgcctaattg atcagatagc tttccatata cagctgttcc
     aatggaaaag gttaacataa aggctgtgtt cacccagttt gtactcgcag gtggtttatt
     aaaatcattt gcaatatcag gtaatgagac gttcaaaacc atttcattta atacgctaaa
     aaaagataaa atgcaaagcc aaattaaaat ttggttgtgt cgtaaattcg attgtgaata
     ggatgtattc acatttcacc ctccaataat gagggcagac gtagtttata gggttaatga
     tacgcttccc tcttttaatt gaaccctgtt acattcatta ttcattacac ttcataatta
     attcctccta aacttgatta aaacatttta ccacatataa actaagtttt aaattcagta
     tttcatcact tatacaacaa tatggcccgt ttgttgaact actctttaat aaaataattt
     ttccgttccc aattccacat tgcaataata gaaaatccat cttcatcggc tttttcgtca
     tcatctgtat gaatcaaatc gccttcttct gtgtcatcaa ggtttaattt tttatgtatt
     tcttttaaca aaccaccata ggagattaac cttttacggt gtaaaccttc ctccaaatca
     gacaaacgtt tcaaattctt ttcttcatca tcggtcataa aatccgtatc ctttacagga
     tattttgcag tttcgtcaat tgccgattgt atatccgatt tatatttatt tttcggtcga
     atcatttgaa cttttacatt tggatcatag tctaatttca ttgccttttt ccaaaattga
     atccattgtt tttgattcac gtagttttct gtattcttaa aataagttgg ttccacacat
     accaatacat gcatgtgctg attataagaa ttatctttat tatttattgt cacttccgtt
     gcacgcataa aaccaacaag atttttatta atttttttat attgcatcat tcggcgaaat
     ccttgagcca tatctgacaa actcttattt aattcttcgc catcataaac atttttaact
     gttaatgtga gaaacaacca acgaactgtt ggcttttgtt taataacttc agcaacaacc
     ttttgtgact gaatgccatg tttcattgct ctcctccagt tgcacattgg acaaagcctg
     gatttacaaa accacactcg atacaacttt ctttcgcctg tttcacgatt ttgtttatac
     tctaatattt cagcacaatc ttttactctt tcagcctttt taaattcaag aatatgcaga
     agttcaaagt aatcaacatt agcgattttc ttttctctcc atggtctcac ttttccactt
     tttgtcttgt ccactaaaac ccttgatttt tcatctgaat aaatgctact attaggacac
     ataatattaa aagaaacccc catctattta gttatttgtt tggtcactta taactttaac
     agatggggtt tttctgtgca accaatttta agggttttca atactttaaa acacatacat
     accaacactt caacgcacct ttcagcaact aaaataaaaa tgacgttatt tctatatgta
     tcaagataag aaagaacaag ttcaaaacca tcaaaaaaag acaccttttc aggtgctttt
     tttattttat aaactcattc cctgatctcg acttcgttct ttttttacct ctcggttatg
     agttagttca aattcgttct ttttaggttc taaatcgtgt ttttcttgga attgtgctgt
     tttatccttt accttgtcta caaacccctt aaaaacgttt ttaaaggctt ttaagccgtc
     tgtacgttcc ttaaggaatt attccttagt gctttctagg ttaatgtcat gataataatg
     gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     aacgacgagc gtgacaccac gatgcctgcc gcaatggcaa caacgttgcg caaactatta
     actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     gcgtcagacc ccttaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct
     tgctctgaaa acgaaaaaac cgccttgcag ggaggttttt cgaaggttct ctgagctacc
     aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca
     gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc
     tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga
     taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac
     tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa
     tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa
     acgcctggta tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt
     cgtgatgctt gtcagggggc ggagcctatg gaaaaacggc tttgccc
//