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ID   PFL64P     preliminary; circular DNA; SYN; 4763 BP.
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AC   X70274; ATCC77213;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Saccharomyces/E.coli phagemid vector pFL64+ - 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   1076-2172
RC   pBR322-8 or pBR322-URA3 from pBR322 & S. cerevisiae URA3 gene
RC   clone 1, clone 2 from pMB9 & yeast S. cerevisiae chromosome
RC   clone 6 from pBR322 & clone 1
RC   pDURA from URA3 gene
RA   Bach M.L., Lacroute F., Botstein D.;
RT   "Evidence for transcriptional regulation of OMP-decarboxylase in
RT   yeast by hybridisation of mRNA to the yesat structural gene cloned
RT   in E. coli";
RL   Proc. Natl. Acad. Sci. U.S.A. 76:386-390(1979).
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RN   [2]
RP   1-4763
RC   pFL64+
RA   Ozier-Kalogeropoulos O.;
RT   ;
RL   Submitted (01-JUN-1993) by:
RL   Ozier-Kalogeropoulos O., CGM, CNRS, 91190 Gif sur Yvette, France.
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RN   [3]
RP   1-4763
RC   plasmid from pUC19 & linkers
RC   pFL26 from plasmid & YEp13, LEU2 gene
RC   pFL34 from plasmid & pBR322-URA3, URA3 gene
RC   pFL35 from plasmid & YRp7, TRP1 gene
RC   pFL36 from pFL26 & CEN6 & ARS
RC   pFL38 from pFL34 & CEN6 & ARS
RC   pFL39 from pFL35 & CEN6 & ARS
RC   pFL46S from pFL26 & yeast 2 micron & linker
RC   pFL44S from pFL34 & yeast 2 micron & linker
RC   pFL45S from pFL35 & yeast 2 micron & linker
RC   pFL46L from pFL26 & pFL1 & linker
RC   pFL44L from pFL34 & pFL1 & linker
RC   pFL45L from pFL35 & pFL1 & linker
RC   pFL59- from pFL39 & pGEM-3Zf-
RC   pFL59+ from pFL44S & pGEM-3Zf+
RC   pFL64- from pFL39 & pGEM-3Zf-
RC   pFL64+ from pFL44S & pGEM-3Zf+
RA   Bonneaud N., Ozier-Kalogeropoulos O., Li G.Y., Labouesse M.,
RA   Minvielle-Sebastia L., Lacroute F.;
RT   "A family of low and high copy replicative, integrative and
RT   single-stranded S. cerevisiae/E. coli shuttle vectors";
RL   Yeast 7:609-615(1991).
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RN   [4]
RP   1-4763
RC   pD30, pD48, pD39, pD40, pD38 from phage f1 & pBR322
RC   pD37 from phage f1 & pBR322
RA   Dotto G.P., Horiuchi K., Zinder N.D.;
RT   "The functional origin of bacteriophage f1 DNA replication";
RL   J. Mol. Biol. 172:507-521(1984).
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CC   pFL64+ was constructed from pGEM-3Zf(+) (Promega) and pUC19
CC   plasmid where two alu I sites were modified. The site 629 was
CC   replaced by a BglII linker and the site 747 by a ClaI site. The
CC   yeast selectable marker has been cloned in the BglII site and the
CC   yeast replication origin in the ClaI site. The 1.4 kb BglI fragment
CC   of the pGEM3Zf+ plasmid bearing the 0.45 kb phage f1 region was
CC   used to replace the coresponding BglI fragment in pFL44.
CC
CC   The order of the major features in this plasmid is:EcoRI/MCS/HindIII -
CC   URA3 - 2 micron ori - pMB1 ori - ampR - f1 ori.
CC   YE-type high copy number shuttle vector permitting visual detection of
CC   recombinants based on alpha complementation. Can be used to produce
CC   ssDNA.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--4.8; BglII--3.7, 1.1; ClaI--4.2, 0.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397139
CC   NM (pFL64+)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(S.cerevisiae)(E.coli)(Saccharomyces)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pUC19)(yeast 2-micron plasmid)(pGEM-3Zf+)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pFL44S remove BglI-BglI 1118bp
FT                   \ 3453..4319..252, 3201bp
FT                   2. pGEM-3Zf+ BglI-BglI 1625bp 3033..3199..1459, f1 ori
FT                   -> pFL64+ 4763bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-BanII-SacI-Asp718-KpnI-AvaI-SmaI-
FT                   XmaI-BamHI-XbaI-
FT                   AccI-HincII-SalI-BspM1-PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SacI-KpnI-SmaI-XmaI-BamHI-
FT                   XbaI-SalI-PstI-SphI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ')"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
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SQ   Sequence 4763 BP; 1259 A; 1110 C; 1107 G; 1287 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tgagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggccac
     gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttaaccaata
     ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     taatgcgccg ctacagggcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcagatc
     tgagcttttt ctttccaatt tttttttttt gtgagtttag tatacatgca tttacttata
     atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc
     tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa
     taataatgtc agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg
     cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat
     ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg
     caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt
     ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac
     cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta
     ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc
     tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc
     cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac
     ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca
     agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag
     tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc cggtttttgt
     tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
     tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     attgaaaagc cagatctgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     cggcgagcgg tatcagcatc gatgcatact ttgtgaacag aaagtgatag cgttgatgat
     tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga acttcggaat
     aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgccatc
     gatgctcact caaaggtcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg gcattgctac
     aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     gcataattct cttactgtca tgccatcggt aagatgcttt tctgtgactg gtgagtactc
     aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     acgggataat agtgtatgac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     gcgtatcacg aggccctttc gtc
//