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ID   PFL35      preliminary; circular DNA; SYN; 3543 BP.
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AC   X70480; ATCC77201;
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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 pFL35 - 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   1-3543
RC   pFL35
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   1-3543
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   pFL35 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.
CC
CC   The order of the major features in this plasmid is:EcoRI/MCS/HindIII -
CC   TRP1 - pMB1 ori - ampR.
CC   YI-type shuttle integration vector permitting visual detection of
CC   recombinants based on alpha complementation.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--3.7; BglII--2.9, 0.85; ClaI--3.7. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397128
CC   NM (pFL35)
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. pUC19 AluI 2686bp 631..631
FT                   BglII linker 6bp agatct
FT                   AluI 2693bp 747..747
FT                   ClaI linker 6bp atcgat
FT                   -> plasmid 2703bp
FT                   1. plasmid BglII 2703bp 630..630
FT                   2. YRp7 AccI 5817bp 1386..1386, TRP1 gene
FT                   fill in
FT                   HindIII 5817bp 1487..1487, TRP1 gene
FT                   mutagenesis
FT                   EcoRI-PstI 823bp 633..1456, TRP1 gene
FT                   fill in:fill in
FT                   BglII linker 6bp agatct:BglII linker 6bp agatct
FT                   -> pFL34 3543bp"
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   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 3543 BP; 919 A; 815 C; 904 G; 905 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 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
//