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ID   PFL45S     preliminary; circular DNA; SYN; 4063 BP.
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AC   X70268; ATCC77206;
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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 plasmid vector pFL45S - 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   1-4063
RC   pFL45S
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   [2]
RP   1-4063
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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CC   pFL45S was constructed from pUC19 plasmid where two alu I sites
CC   were modified. The site 629 was replaced by a BglII linker and the
CC   site 747 by a ClaI site. The yeast selectable marker has been
CC   cloned in the BglII site and the 2 micron 0.5 Kb FspI fragment
CC   containing its replication origin has been cloned at the ClaI site.
CC
CC   The order of the major features in this plasmid is:EcoRI/MCS/HindIII -
CC   TRP1 - 2 micron ori - pMB1 ori - ampR.
CC   YE-type high copy number shuttle vector permitting visual detection of
CC   recombinants based on alpha complementation.
CC   The loss per generation (percentage having lost all the plasmids after
CC   20 h of non-selective growth) is 11.0.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--4.2; BglII--3.4, 0.8; ClaI--3.7, 0.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397133
CC   NM (pFL45S)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
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)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pFL35 ClaI 3543bp 1600..1600
FT                   2. yeast 2 micron XbaI 6318bp 3946..3946
FT                   fill in
FT                   FspI-FspI 509bp 3516..4025, ori
FT                   ClaI linker 6bp atcgat:ClaI linker 6bp atcgat
FT                   -> pFL45S 4063bp"
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                   AccI-HincII-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 yeast 2 micron"
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 TRP1 gene"
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SQ   Sequence 4063 BP; 1073 A; 892 C; 1021 G; 1077 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
     cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     gctttccagt cgggaaacct gtcgtgccag cagatctgaa ttaattcggt cgaaaaaaga
     aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt atatataccg
     tgattaagca cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg
     gtccattggt gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt
     ccgatgctga cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg
     ttattgcaag gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga
     aatacttggt tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg
     attacggcat tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt
     tcctcggttt gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca
     gtgcagcttc acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga
     caggtgaact tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg
     aaagtttaca ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta
     gattaaatgg cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag
     actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt
     atttatttaa gtattgtttg tgcacttgcc 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 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     tgctcttgcc cggcgtcaat acgggataat accgcgccac 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
//