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ID   YEP352     preliminary; circular DNA; SYN; 5181 BP.
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AC   L14758; ATCC37673;
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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 YEp352 - 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   1-5181
RC   YEp351 from pUC18 & pC4B, yeast 2 micron/LEU2 gene
RC   YEp352 from pUC18 & yeast 2 micron & yeast URA3 gene
RC   YIp351 from YEp351
RC   YIp352 from YEp352
RA   Hill J.E., Myers A.M., Koerner T.J., Tzagoloff A.;
RT   "Yeast/E. coli shuttle vectors with multiple unique restriction
RT   sites";
RL   Yeast 2:163-167(1986).
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RN   [2]
RC   YEp352 from pUC18 & yeast 2 micron & yeast URA3 gene
RA   ;
RT   ;
RL   Unpublished (1988).
RL   U.S. Patent No. 4,794,175 dated Dec. 27, 1988.
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CC   Constructed by filling in and blunt-end ligating pUC18 linearized at
CC   NdeI, a 1.4 kb RsaI fragment of the 2 micron circle, and a 1.1 kb
CC   HindIII/SmaI fragment containing URA3. A XbaI site in 2u and a PstI
CC   site in URA3 were removed. [1]
CC   The yeast genomic sequence is in nt 286-1383 (where the A of the ATG
CC   of lacZ' is number 1), the 2 micron sequence goes from nt 1384-2768,
CC   & pUC18 includes nt 2769-285. [2]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.3; XbaI--5.3; PstI--5.3; HindIII--5.3. (ATCC staff)
CC   This is a E.coli/yeast episomal shuttle vector with multiple
CC   restriction sites for cloning. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 295271
CC   NM (YEp352)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli RR1)(S.cerevisiae)(E.coli)(Saccharomyces)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pBR322)(2micron)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC18 HindIII 2686bp 400..400
FT                   2. YEp24 HindIII-HindIII 1166bp 2273..3439, URA3 gene
FT                   -> pUC18::URA3 3852bp
FT                   1. pUC18 NdeI 2686bp 185..185, MCS
FT                   Klenow
FT                   2. yeast 2 micron RsaI-RsaI 1416bp 526..1942,
FT                   \ ori/may be 1130..2452
FT                   3. pUC18::URA3 HindIII-SmaI 1108bp 2273..3381,
FT                   \ yeast URA3 gene
FT                   Klenow:Klenow
FT                   -> plasmid 5200bp
FT                   1. plasmid XbaI 5200bp, yeast 2 micron 704..704
FT                   Klenow
FT                   PstI, URA3 gene/YEp24 2482..2482
FT                   S1 nuclease, remove 20bp
FT                   -> YEp352 5181bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-KpnI-SmaI-BamHI-XbaI-SalI-
FT                   PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SacI-KpnI-SmaI-BamHI-XbaI-
FT                   SalI-PstI-SphI-HindIII-Narl-
FT                   StuI-ApaI-AsuII-NcoI-NdeI-AatII-HgiEII"
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 URA3 gene"
FT   misc_feature    1..285
FT                   /note="from Escherichia coli"
FT   misc_feature    286..2768
FT                   /note="from Saccharomyces cerevisiae"
FT   misc_feature    2769..5181
FT                   /note="from Escherichia coli"
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SQ   Sequence 5181 BP; 1377 A; 1201 C; 1130 G; 1473 T; 0 other;
     atgaccatga ttacgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg
     catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     acccaactta atcgccttgc agcacatccc cccttcgcca gctggcgtaa tagcgaagag
     gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata
     taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcggttttt gttctgtgca
     gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     gctcttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     ttcctatact ttctagctag agaataggaa cttcggaata ggaacttcaa agcgtttccg
     aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc
     gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt
     gtttatgctt aaatgcgtta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     aaccgcctct ccccgcgcgt tggccgattc attaatccag ctggcacgac aggtttcccg
     actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac
     cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     aatttcacac aggaaacagc t
//