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ID PFL46L preliminary; circular DNA; SYN; 7822 BP.
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AC X70269; ATCC77210;
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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 pFL46L - 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 1-7822
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 ()
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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. 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"
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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
//