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ID   PFL46S     preliminary; circular DNA; SYN; 6078 BP.
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AC   X70270; ATCC77207;
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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 pFL46S - 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   1-6078
RC   pFL46S
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   1-6078
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   pFL46S 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   LEU2 - 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 15.5.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--6.1; BglII--3.3, 2.8; ClaI--3.7, 1.9, 0.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397135
CC   NM (pFL46S)
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. pFL26 ClaI 5558bp 1694..1694
FT                   2. yeast 2 micron 6318bp 3946..3946
FT                   fill in
FT                   FspI-FspI 509bp 3516..4025, ori
FT                   ClaI linker 6bp atcgat:ClaI linker 6bp atcgat
FT                   -> pFL46S 6078bp"
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 SacI-SmaI-XmaI-BamHI-XbaI-PstI-
FT                   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 LEU2 gene"
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SQ   Sequence 6078 BP; 1740 A; 1295 C; 1368 G; 1675 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 cagatctatt acattatggg tggtatgttg
     gaataaaaat caactatcat ctactaacta gtatttacgt tactagtata ttatcatata
     cggtgttaga agatgacgca aatgatgaga aatagtcatc taaattagtg gaagctgaaa
     cgcaaggatt gataatgtaa taggatcaat gaatattaac atataaaatg atgataataa
     tatttataga attgtgtaga attgcagatt cccttttatg gattcctaaa tcctcgagga
     gaacttctag tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca
     attacatcaa aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg
     tgttcaaaaa cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt
     gtttggccga gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg
     ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt
     cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga
     aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg
     gagaaaaagg aaaggtgaga gccgcggaac cggcttttca tatagaatag agaagcgttc
     atgactaaat gcttgcatca caatacttga agttgacaat attatttaag gacctattgt
     tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt
     cagcaatata tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga
     tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc
     ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg
     gtggtgctgc tatcgatgct acaggtgtcc cacttccaga tgaggcgctg gaagcctcca
     agaaggttga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg
     ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa
     gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg
     ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga
     gaaaggaaga cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag
     tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta
     tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg
     aggaaaccat caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg
     ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca
     tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc
     catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat
     gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt
     ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag
     atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca
     gtaccacgga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag
     attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg
     acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac
     atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat
     actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga
     ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt
     tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg
     acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga
     aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagt cgacctcctc
     ttcagtacta aatctaccaa taccaaatct gatggaagaa tgggctaatg catcatcctt
     acccagcgca tgtaaaacat aagaaggttc tagggaagca gatgtacagg ctgaacccga
     ggataatgcg atatccctta gtgccatcaa taaagattct ccttccacgt aggcgaaaga
     aacgttaaca caccctggat aacgatgatc tggagatccg ttcaacgtgg tatgttcagc
     ggataataga cctttgacta atttatcgga tagtcttttg atgtgagctt ggtcgttgtc
     aaattctttc ttcatcaatc tcgcagcttc accaaatccc gctaccaatg ggggggccaa
     agtaccagat ctgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     agcggtatca gcatcgatgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt
     caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ggaataggaa
     cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg ccatcgatgc
     tcactcaaag gtcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     tcacgaggcc ctttcgtc
//