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ID PFL44L preliminary; circular DNA; SYN; 6063 BP.
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AC X70484; ATCC77208;
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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 pFL44L - 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 644-1740
RC pBR322-8 or pBR322-URA3 from pBR322 & S. cerevisiae URA3 gene
RC clone 1, clone 2 from pMB9 & yeast S. cerevisiae chromosome
RC clone 6 from pBR322 & clone 1
RC pDURA from URA3 gene
RA Bach M.L., Lacroute F., Botstein D.;
RT "Evidence for transcriptional regulation of OMP-decarboxylase in
RT yeast by hybridisation of mRNA to the yesat structural gene cloned
RT in E. coli";
RL Proc. Natl. Acad. Sci. U.S.A. 76:386-390(1979).
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RN [2]
RP 1-6063
RC pFL44L
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 [3]
RP 1-6063
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 pFL44L 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 URA3 - 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 4.0.
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--6.1; BglII--5.0, 1.1; ClaI--3.9, 2.25. (ATCC staff)
CC Medium is 1227 LB plus ampicillin.
CC NCBI gi: 312626
CC NM (pFL44L)
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. pFL34 ClaI 3799bp 1856..1856
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 6000bp
FT 1. plasmid EcoRI 6000bp 2..2
FT NotI linker 8bp gcggccgc
FT -> pFL44L 6063bp"
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 URA3 gene"
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SQ Sequence 6063 BP; 1607 A; 1385 C; 1307 G; 1764 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 agctttttct
ttccaatttt ttttttttgt gagtttagta tacatgcatt tacttataat acagtttttt
agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
ccttatatgt agctttcgac atgatttatc ttcgtttccg gtttttgttc tgtgcagttg
ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcca
gatctgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
tcagcatcga tgaattccac ggactataga ctatactagt atactccgtc tactgtacga
tacacttccg ctcaggtcct tgtcctttaa cgaggcctta ccactctttt gttactctat
tgatccagct cagcaaaggc agtgtgatct aagattctat cttcgcgatg tagtaaaact
agctagaccg agaaagagac tagaaatgca aaaggcactt ctacaatggc tgccatcatt
attatccgat gtgacgctgc agcttctcaa tgatattcga atacgctttg aggagataca
gcctaatatc cgacaaactg ttttacagat ttacgatcgt acttgttacc catcattgaa
ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata
ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac
tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt
ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga
acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagagaata
ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg aaaatgcaac
gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt gttgcctgta
tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc gtacttatat
gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta tcccattcca
tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat gctgccactc
ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg gatcatatgc
atagtaccga gaaactagtg cgaagtagtg atcaggtatt gctgttatct gatgagtata
cgttgtcctg gccacggcag aagcacgctt atcgctccaa tttcccacaa cattagtcaa
ctccgttagg cccttcattg aaagaaatga ggtcatcaaa tgtcttccaa tgtgagattt
tgggccattt tttatagcaa agattgaata aggcgcattt ttcttcaaag ctttattgta
cgatctgact aagttatctt ttaataattg gtattcctgt ttattgcttg aagaattgcc
ggtcctattt actcgtttta ggactggttc agaattcatc gatgctcact caaaggtcgg
taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
gtc
//