ID   YEP352     preliminary; circular DNA; SYN; 5181 BP.

XX

AC   L14758; ATCC37673;

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 YEp352 - complete.

XX

KW   cloning vector.

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OS   Cloning vector

OC   Artificial sequences; Cloning vehicles.

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RN   [1]

RP   1-5181

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RL   Unpublished (1988).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"

XX

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

//