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ID   PFL44L     preliminary; circular DNA; SYN; 6063 BP.
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AC   X70484; ATCC77208;
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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 pFL44L - 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   644-1740
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   1-6063
RC   pFL44L
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   1-6063
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   pFL44L 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 2.2 kb EcoRI fragment
CC   containing ORI and STB gene has been cloned at the ClaI site.
CC
CC   The order of the major features in this plasmid is:NotI/MCS/HindIII -
CC   URA3 - STB -2 micron ori - pMB1 ori - ampR.
CC   YE-type high copy number shuttle vector.
CC   Shows increased stability because of the presence of STB.  The loss
CC   per generation (percentage having lost all the plasmids after 20 h of
CC   non-selective growth) is 4.0.
CC   Contrary to published data, this vector does not permit visual
CC   detection of recombinants using the assay for alpha complementation.
CC   (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--6.1; BglII--5.0, 1.1; ClaI--3.9, 2.25. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 312626
CC   NM (pFL44L)
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. yeast 2 micron remove, 6318bp-
FT                   -> pFL1 6000bp
FT                   1. pFL34 ClaI 3799bp 1856..1856
FT                   2. pFL1 EcoRI-EcoRI 2200bp, yeast 2 micron 2..-..2408
FT                   \ yeast 2 micron ori/STB gene
FT                   fill in:fill in
FT                   ClaI linker 6bp atcgat:ClaI linker 6bp atcgat
FT                   -> plasmid 6000bp
FT                   1. plasmid EcoRI 6000bp 2..2
FT                   NotI linker 8bp gcggccgc
FT                   -> pFL44L 6063bp"
FT   misc_binding    0..0
FT                   /note="MCS NotI-EagI-BanII-SacI-Asp718-KpnI-AvaI-
FT                   SmaI-XmaI-BamHI-
FT                   XbaI-AccI-HincII-SalI-BspM1-PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique NotI-SacI-KpnI-SmaI-XmaI-BamHI-
FT                   SalI-SphI"
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   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 6063 BP; 1607 A; 1385 C; 1307 G; 1764 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 gcggccgcaa ttcgagctcg
     gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta atcatggtca
     tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcagatctg agctttttct
     ttccaatttt ttttttttgt gagtttagta tacatgcatt tacttataat acagtttttt
     agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
     cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
     atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
     aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
     ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     ccttatatgt agctttcgac atgatttatc ttcgtttccg gtttttgttc tgtgcagttg
     ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcca
     gatctgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     tcagcatcga tgaattccac ggactataga ctatactagt atactccgtc tactgtacga
     tacacttccg ctcaggtcct tgtcctttaa cgaggcctta ccactctttt gttactctat
     tgatccagct cagcaaaggc agtgtgatct aagattctat cttcgcgatg tagtaaaact
     agctagaccg agaaagagac tagaaatgca aaaggcactt ctacaatggc tgccatcatt
     attatccgat gtgacgctgc agcttctcaa tgatattcga atacgctttg aggagataca
     gcctaatatc cgacaaactg ttttacagat ttacgatcgt acttgttacc catcattgaa
     ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata
     ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac
     tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt
     ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga
     acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
     ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac
     gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta
     tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat
     gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca
     tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc
     ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatatgc
     atagtaccga gaaactagtg cgaagtagtg atcaggtatt gctgttatct gatgagtata
     cgttgtcctg gccacggcag aagcacgctt atcgctccaa tttcccacaa cattagtcaa
     ctccgttagg cccttcattg aaagaaatga ggtcatcaaa tgtcttccaa tgtgagattt
     tgggccattt tttatagcaa agattgaata aggcgcattt ttcttcaaag ctttattgta
     cgatctgact aagttatctt ttaataattg gtattcctgt ttattgcttg aagaattgcc
     ggtcctattt actcgtttta ggactggttc agaattcatc 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
//