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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
//