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ID   PFL46L     preliminary; circular DNA; SYN; 7822 BP.
XX
AC   X70269; ATCC77210;
XX
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 pFL46L - complete.
XX
KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-7822
RC   pFL46L
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   1-7822
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   pFL46L 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   LEU2 - 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 havaing lost all the plasmids after 20 h of
CC   non-selective growth) is 6.3.
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--7.8; BglII--4.9, 2.9; ClaI--3.7, 2.3, 1.9. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 397134
CC   NM (pFL46L)
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 ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. yeast 2 micron remove, 6318bp-
FT                   -> pFL1 6000bp
FT                   1. pFL26 ClaI 5558bp 1694..1694
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 7800bp
FT                   1. plasmid EcoRI 7800bp 2..2
FT                   NotI linker 8bp gcggccgc
FT                   -> pFL46L 7822bp"
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-SmaI-XmaI-BamHI-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 LEU2 gene"
XX
SQ   Sequence 7822 BP; 2222 A; 1664 C; 1691 G; 2245 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 agcagatcta ttacattatg
     ggtggtatgt tggaataaaa atcaactatc atctactaac tagtatttac gttactagta
     tattatcata tacggtgtta gaagatgacg caaatgatga gaaatagtca tctaaattag
     tggaagctga aacgcaagga ttgataatgt aataggatca atgaatatta acatataaaa
     tgatgataat aatatttata gaattgtgta gaattgcaga ttccctttta tggattccta
     aatcctcgag gagaacttct agtatatcta catacctaat attattgcct tattaaaaat
     ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc
     cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac
     aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg
     cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac
     cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat
     tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa
     ctgaaaaatt gggagaaaaa ggaaaggtga gagccgcgga accggctttt catatagaat
     agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta
     aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta
     ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt ctaatgtctg
     cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc acagccgaag
     ccattaaggt tcttaaagct atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa
     atcatttaat tggtggtgct gctatcgatg ctacaggtgt cccacttcca gatgaggcgc
     tggaagcctc caagaaggtt gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg
     gtaccggtag tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt
     acgccaactt aagaccatgt aactttgcat ccgactctct tttagactta tctccaatca
     agccacaatt tgctaaaggt actgacttcg ttgttgtcag agaattagtg ggaggtattt
     actttggtaa gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt gaacaataca
     ccgttccaga agtgcaaaga atcacaagaa tggccgcttt catggcccta caacatgagc
     caccattgcc tatttggtcc ttggataaag ctaatgtttt ggcctcttca agattatgga
     gaaaaactgt ggaggaaacc atcaagaacg aattccctac attgaaggtt caacatcaat
     tgattgattc tgccgccatg atcctagtta agaacccaac ccacctaaat ggtattataa
     tcaccagcaa catgtttggt gatatcatct ccgatgaagc ctccgttatc ccaggttcct
     tgggtttgtt gccatctgcg tccttggcct ctttgccaga caagaacacc gcatttggtt
     tgtacgaacc atgccacggt tctgctccag atttgccaaa gaataaggtc aaccctatcg
     ccactatctt gtctgctgca atgatgttga aattgtcatt gaacttgcct gaagaaggta
     aggccattga agatgcagtt aaaaaggttt tggatgcagg tatcagaact ggtgatttag
     gtggttccaa cagtaccacg gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc
     ttgcttaaaa agattctctt tttttatgat atttgtacat aaactttata aatgaaattc
     ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaaactata
     tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag gaatacaggt
     aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa
     tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta
     tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa
     aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc
     ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct taacgacgta
     gtcgacctcc tcttcagtac taaatctacc aataccaaat ctgatggaag aatgggctaa
     tgcatcatcc ttacccagcg catgtaaaac ataagaaggt tctagggaag cagatgtaca
     ggctgaaccc gaggataatg cgatatccct tagtgccatc aataaagatt ctccttccac
     gtaggcgaaa gaaacgttaa cacaccctgg ataacgatga tctggagatc cgttcaacgt
     ggtatgttca gcggataata gacctttgac taatttatcg gatagtcttt tgatgtgagc
     ttggtcgttg tcaaattctt tcttcatcaa tctcgcagct tcaccaaatc ccgctaccaa
     tgggggggcc aaagtaccag atctgctgca ttaatgaatc ggccaacgcg cggggagagg
     cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     tcggctgcgg cgagcggtat cagcatcgat gaattccacg gactatagac tatactagta
     tactccgtct actgtacgat acacttccgc tcaggtcctt gtcctttaac gaggccttac
     cactcttttg ttactctatt gatccagctc agcaaaggca gtgtgatcta agattctatc
     ttcgcgatgt agtaaaacta gctagaccga gaaagagact agaaatgcaa aaggcacttc
     tacaatggct gccatcatta ttatccgatg tgacgctgca gcttctcaat gatattcgaa
     tacgctttga ggagatacag cctaatatcc gacaaactgt tttacagatt tacgatcgta
     cttgttaccc atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata
     gtatatttga acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta
     tttcggttcc tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc
     cggttcattt tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat
     ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt
     ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     ttgatattgg atcatatgca tagtaccgag aaactagtgc gaagtagtga tcaggtattg
     ctgttatctg atgagtatac gttgtcctgg ccacggcaga agcacgctta tcgctccaat
     ttcccacaac attagtcaac tccgttaggc ccttcattga aagaaatgag gtcatcaaat
     gtcttccaat gtgagatttt gggccatttt ttatagcaaa gattgaataa ggcgcatttt
     tcttcaaagc tttattgtac gatctgacta agttatcttt taataattgg tattcctgtt
     tattgcttga agaattgccg gtcctattta ctcgttttag gactggttca gaattcatcg
     atgctcactc aaaggtcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     cgtatcacga ggccctttcg tc
//