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ID   PHY300PLK  preliminary; circular DNA; SYN; 4870 BP.
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AC   D00054; N00054;
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DT   19-SEP-1987 (Rel. 2, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Bacillus/E.coli plasmid vector pHY300PLK - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-4870
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).
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RN   [2]
RC   pHY385 from pHY460
RC   pHY340 from pHY385
RC   pHY339 from pHY340
RC   pHY336 from pHY339 & linker
RC   pHY300PLK from pHY336 & piAN7
RA   Ishiwa H., Shibahara H.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis.
RT   II. Plasmid pHY300PLK, a multipurpose cloning vector with a
RT   polylinker, derived from pHY460";
RL   Jpn. J. Genet. 60:235-243(1985).
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RN   [3]
RC   pHY460 from pACYC177 & pAMalpha1
RA   Ishiwa H., Tsuchida N.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis.
RT   I. Construction and characterization of plasmid pHY460 with twelve
RT   unique cloning sites";
RL   Gene 32:129-134(1984).
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RN   [4]
RC   pHY300-7PLK from pHY300PLK & pHY340
RC   pHY300-1PLK from pHY300-7PLK & linker
RC   pHY230PLK from pHY300-1PLK & linker
RC   pHY163PLK from pHY230PLK
RA   Ishiwa H., Shibahara H.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis.
RT   III. Nucleotide sequence analysis of tetracycline resistance gene
RT   of pAMalpha1 and ori-177";
RL   Jpn. J. Genet. 60:485-498(1985).
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CC   PHY300PLK 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. This shuttle vector for
CC   E.coli and B.subtilis contains an RNA primer gene for ori-177; the
CC   R-Tc gene; the R-Ap gene; Rep-alpha-1 gene, which is the plasmid
CC   replication gene; two replication origins, one for E.coli and the
CC   other for B. subtilis; and a polylinker. Twenty-three unique
CC   recognition restriction endonuclease sites have been found in
CC   pHY300PLK.
CC   NM (pHY300PLK)
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)
CC   BR (pHY320PLK)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pACYC177, amp/kan gene
FT                   2. pAMalpha1, tet gene
FT                   -> pHY460 7000bp
FT                   1. pHY460 remove between pAMalpha1 ori and amp
FT                   spontaneous deletion
FT                   -> pHY385 5900bp [no XbaI, no AvaI, no BamHI]
FT                   1. pHY385 remove HaeII-HaeII, between amp and tet
FT                   -> pHY340 5200bp [unique HaeII]
FT                   1. pHY340 EcoRI, between pAMalpha1 ori and amp
FT                   T4 DNA polymerase
FT                   -> pHY339 5200bp [unique EcoRI]
FT                   1. pHY339 SacII
FT                   T4 DNA polymerase
FT                   HindIII linker 6bp aagctt
FT                   -> pHY336 5200bp
FT                   1. pHY336 remove small EcoRI-HindIII, 5' to tet
FT                   2. piAN7 EcoRI-HindIII, MCS
FT                   -> pHY300PLK 4870bp"
FT   misc_feature    1..42
FT                   /note="MCS HindIII-XbaI-Bgl-Pst-AccI-HincII-SalI-
FT                   BamHI-AvaI-SmaI-EcoRI"
FT   CDS             complement(100..1476)
FT                   /note="ANT E. coli tetracycline resistance gene (tet);
FT                   open reading frame (ORF)"
FT   CDS             complement(1683..2687)
FT                   /note="GEN E. coli Rep-alpha-1 gene ORF"
FT   CDS             3289..4149
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp) ORF"
FT   CDS             4350..4860
FT                   /note="GEN E. coli RNA primer gene ori-177"
FT   rep_origin      0..0
FT                   /note="ORI E. coli p15a"
FT   rep_origin      0..0
FT                   /note="ORI Bacillus pUB110"
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SQ   Sequence 4870 BP; 1474 A; 1026 C; 867 G; 1503 T; 0 other;
     aagcttctag agatctgcag gtcgacggat ccccgggaat tcctgttata aaaaaaggat
     caattttgaa ctctctccca aagttgatcc cttaacgatt tagaaatccc tttgagaatg
     tttatataca ttcaaggtaa ccagccaact aatgacaatg attcctgaaa aaagtaataa
     caaattacta tacagataag ttgactgatc aacttccata ggtaacaacc tttgatcaag
     taagggtatg gataataaac cacctacaat tgcaatacct gttccctctg ataaaaagct
     ggtaaagtta agcaaactca ttccagcacc agcttcctgc tgtttcaagc tacttgaaac
     aattgttgat ataactgttt tggtgaacga aagcccacct aaaacaaata cgattataat
     tgtcatgaac catgatgttg tttctaaaag aaaggaagca gttaaaaagc taacagaaag
     aaatgtaact ccgatgttta acacgtataa aggacctctt ctatcaacaa gtatcccacc
     aatgtagccg aaaataatga cactcattgt tccagggaaa ataattacac ttccgatttc
     ggcagtactt agctggtgaa catctttcat catataagga accatagaga caaaccctgc
     tactgttcca aatataattc ccccacaaag aactccaatc ataaaaggta tatttttccc
     taatccggga tcaacaaaag gatctgttac tttcctgata tgttttacaa atatcaggaa
     tgacagcacg ctaacgataa gaaaagaaat gctatatgat gttgtaaaca acataaaaaa
     tacaatgcct acagacatta gtataattcc tttgatatca aaatgacctt ttatccttac
     ttctttcttt aataatttca taagaaacgg aacagtgata attgttatca taggaatgag
     tagaagatag gaccaatgaa tataatgggc tatcattcca ccaatcgctg gaccgactcc
     ttctcccatg gctactatcg atccaataag accaaatgct ttacccctat tttcctttgg
     aatatagcgc gcaactacaa ccattacgag tgctggaaat gcagctgcac cagccccttg
     aataaaacga gccataataa gtaaggaaaa gaaagaatgg ccaacaaacc caattaccga
     cccgaaacaa tttattataa ttccaaatag gagtaacctt ttgatgccta attgatcaga
     tagctttcca tatacagctg ttccaatgga aaaggttaac ataaaggctg tgttcaccca
     gtttgtactc gcaggtggtt tattaaaatc atttgcaata tcaggtaatg agacgttcaa
     aaccatttca tttaatacgc taaaaaaaga taaaatgcaa agccaaatta aaatttggtt
     gtgtcgtaaa ttcgattgtg aataggatgt attcacattt caccctccaa taatgagggc
     agacgtagtt tatagggtta atgatacgct tccctctttt aattgaaccc tgttacattc
     attattcatt acacttcata attaattcct cctaaacttg attaaaacat tttaccacat
     ataaactaag ttttaaattc agtatttcat cacttataca acaatatggc ccgtttgttg
     aactactctt taataaaata atttttccgt tcccaattcc acattgcaat aatagaaaat
     ccatcttcat cggctttttc gtcatcatct gtatgaatca aatcgccttc ttctgtgtca
     tcaaggttta attttttatg tatttctttt aacaaaccac cataggagat taacctttta
     cggtgtaaac cttcctccaa atcagacaaa cgtttcaaat tcttttcttc atcatcggtc
     ataaaatccg tatcctttac aggatatttt gcagtttcgt caattgccga ttgtatatcc
     gatttatatt tatttttcgg tcgaatcatt tgaactttta catttggatc atagtctaat
     ttcattgcct ttttccaaaa ttgaatccat tgtttttgat tcacgtagtt ttctgtattc
     ttaaaataag ttggttccac acataccaat acatgcatgt gctgattata agaattatct
     ttattattta ttgtcacttc cgttgcacgc ataaaaccaa caagattttt attaattttt
     ttatattgca tcattcggcg aaatccttga gccatatctg acaaactctt atttaattct
     tcgccatcat aaacattttt aactgttaat gtgagaaaca accaacgaac tgttggcttt
     tgtttaataa cttcagcaac aaccttttgt gactgaatgc catgtttcat tgctctcctc
     cagttgcaca ttggacaaag cctggattta caaaaccaca ctcgatacaa ctttctttcg
     cctgtttcac gattttgttt atactctaat atttcagcac aatcttttac tctttcagcc
     tttttaaatt caagaatatg cagaagttca aagtaatcaa cattagcgat tttcttttct
     ctccatggtc tcacttttcc actttttgtc ttgtccacta aaacccttga tttttcatct
     gaataaatgc tactattagg acacataata ttaaaagaaa cccccatcta tttagttatt
     tgtttggtca cttataactt taacagatgg ggtttttctg tgcaaccaat tttaagggtt
     ttccaatact ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat
     aaaaatgacg ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa
     aaaagacacc ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc
     gttctttttt tacctctcgg ttatgagtta gttcaaattc gttcttttta ggttctaaat
     cgtgtttttc ttggaattgt gctgttttat cctttacctt gtctacaaac cccttaaaaa
     cgtttttaaa ggcttttaag cgtctgtacg ttccttaagg aattattcct tagtgctttc
     taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     gtccgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc ttcttgagat
     cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggaggt
     ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tgcaggagcg
     cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     accactgatt tgagcgtcag atttcgtgat gcttgtcagg gggcggagcc tatggaaaaa
     cgctttgccc
//