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ID   PFL45L     preliminary; circular DNA; SYN; 5807 BP.
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AC   X70267; ATCC77209;
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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 pFL45L - 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   1-5807
RC   pFL45L
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   1-5807
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   pFL45L 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   TRP1 - 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 4.8.
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--6.2; BglII--5.2, 0.85; ClaI--3.7, 2.25. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397132
CC   NM (pFL45L)
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. pFL35 ClaI 3543bp 1600..1600
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 5800bp
FT                   1. plasmid EcoRI 5800bp 2..2
FT                   NotI linker 8bp gcggccgc
FT                   -> pFL45L 5807bp"
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 TRP1 gene"
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SQ   Sequence 5807 BP; 1555 A; 1261 C; 1344 G; 1647 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 aattaattcg
     gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt gagcacgtga
     gtatatatac cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     aagagagccc cgaaagttta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     ttactgagta gtatttattt aagtattgtt tgtgcacttg cccagatctg ctgcattaat
     gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagca tcgatgaatt
     ccacggacta tagactatac tagtatactc cgtctactgt acgatacact tccgctcagg
     tccttgtcct ttaacgaggc cttaccactc ttttgttact ctattgatcc agctcagcaa
     aggcagtgtg atctaagatt ctatcttcgc gatgtagtaa aactagctag accgagaaag
     agactagaaa tgcaaaaggc acttctacaa tggctgccat cattattatc cgatgtgacg
     ctgcagcttc tcaatgatat tcgaatacgc tttgaggaga tacagcctaa tatccgacaa
     actgttttac agatttacga tcgtacttgt tacccatcat tgaattttga acatccgaac
     ctgggagttt tccctgaaac agatagtata tttgaacctg tataataata tatagtctag
     cgctttacgg aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca
     taggtaatct tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa
     tagcatatct ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact tcggaatagg
     aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag
     gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
     cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
     tcatccttca atgctatcat ttcctttgat attggatcat atgcatagta ccgagaaact
     agtgcgaagt agtgatcagg tattgctgtt atctgatgag tatacgttgt cctggccacg
     gcagaagcac gcttatcgct ccaatttccc acaacattag tcaactccgt taggcccttc
     attgaaagaa atgaggtcat caaatgtctt ccaatgtgag attttgggcc attttttata
     gcaaagattg aataaggcgc atttttcttc aaagctttat tgtacgatct gactaagtta
     tcttttaata attggtattc ctgtttattg cttgaagaat tgccggtcct atttactcgt
     tttaggactg gttcagaatt catcgatgct cactcaaagg tcggtaatac ggttatccac
     agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//