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ID   PRS425     preliminary; circular DNA; SYN; 6849 BP.
XX
AC   U03452; ATCC77106;
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 phagemid vector pRS425 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-6849
RC   plasmid from YEp24 & pRSS56
RC   [pBR322/SC4128 from pBR322 & yeast]
RC   pJS92 from plasmid & pBR322/SC4128
RC   pRS323 from pJS92 & pRS303
RC   pRS324 from pJS92 & pRS304
RC   pRS325 from pJS92 & pRS305
RC   pRS326 from pJS92 & pRS306
RC   pRS423 from pRS323 & pBluescript II SK+
RC   pRS424 from pRS324 & pBluescript II SK+
RC   pRS425 from pRS325 & pBluescript II SK+
RC   pRS426 from pRS326 & pBluescript II SK+
RA   Christianson T.W., Sikorski R.S., Dante M., Shero J.H., Hieter P.;
RT   "Multifunctional yeast high-copy-number shuttle vectors";
RL   Gene 110:119-122(1992).
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RN   [2]
RP   1-6849
RC   pRSS56 from pBluescript KS+ & BlueScribe
RC   pRS303 from pRSS56 & HIS3 gene
RC   pRS304 from pRSS56 & TRP1 gene
RC   pRS305 from pRSS56 & LEU2 gene
RC   pRS306 from pRSS56 & URA3 gene
RC   pUC19-CEN6:32 from pUC19 & CEN6
RC   pRSS83 from pUC19-CEN6:32 & pAB9, ARS
RC   pRSS84 from pRSS83
RC   pRS313 from pRS303 & pRSS84
RC   pRS314 from pRS304 & pRSS84
RC   pRS315 from pRS305 & pRSS84
RC   pRS316 from pRS306 & pRSS84
RC   pRSS93 from pRSS84 & pYCF5
RA   Sikorski R.S., Hieter P.;
RT   "A system of shuttle vectors and yeast host strains designed for
RT   efficient manipulation of DNA in Saccharomyces cerevisiae";
RL   Genetics 122:19-27(1989).
XX
RN   [3]
RC   pRS425
RA   Stillman D.J.;
RT   ;
RL   Submitted (10-NOV-1993) by:
RL   Stillman D.J., Dept. of Cellular, Viral and Molecular Biology,
RL   University of Utah Medical Center, Salt Lake City, UT 84132 USA.
XX
CC   The order of the major features in this plasmid is:ori(f1) - lacZ - T7
CC   promoter - MCS (KpnI-SacI) - T3 promoter - lacI - ori(pMB1) - ampR -
CC   ori (2 micron) - LEU2.
CC   Contains the REP3 and FRT sequences necessary for high copy
CC   propagation in yeast.
CC   In S. cerevisiae, the copy number is about 20 per haploid cell.  In
CC   non-selective growth, plasmids are lost through mitotic segregation at
CC   rates in the range of 4.6 + or - 0.6% of progeny per doubling.
CC   YE-type shuttle vector encoding the beta-galactosidase alpha peptide,
CC   permitting blue-white visual detection. Can be used to produce ssDNA.
CC   Contains promoters for in vitro RNA synthesis and priming sites useful
CC   for sequencing.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--6.8; EcoRI--4.8, 2.1; XbaI--3.6, 3.2; PvuII--6.4, 0.5. (ATCC
CC   staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416323
CC   NM (pRS425)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli XL1-Blue)(Saccharomyces cerevisiae)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR (pRS424)(pRS423)(pRS426)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. YEp24 NdeI-HpaI 1346bp 341..1687,
FT                   \ yeast 2 micron ori [1345bp]
FT                   :Klenow
FT                   2. pRSS56 AatII 2998bp 3140..3140
FT                   T4 DNA polymerase
FT                   -> plasmid 4344bp [unique NdeI]
FT                   1. pBR322 4361bp
FT                   2. yeast 1800bp, TRP1 gene
FT                   -> pBR322/SC4128 6200bp
FT                   1. plasmid NdeI 4344bp 185..185
FT                   Klenow
FT                   2. pBR322/SC4128 BglII-BglII 1800bp, TRP1 gene
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pJS92 6250bp
FT                   1. pJS92 remove DraII-DraII 1800bp, TRP1 gene/4445bp
FT                   2. pRS305 DraII-DraII 1345bp 1876..3221, LEU2 gene
FT                   -> pRS325 6849bp
FT                   1. pRS325 remove PvuI-PvuI 1913bp 2975..4888,
FT                   \ MCS/4936bp
FT                   2. pBluescript II SK+ PvuI-PvuI 1916bp 501..2417, MCS
FT                   -> pRS425 6849bp"
FT   misc_binding    0..0
FT                   /note="MCS SacI-BstXI-SacII-EagI-NotI-XbaI-SpeI-
FT                   BamHI-SmaI-PstI-
FT                   EcoRI-EcoRV-HindIII-ClaI-SalI-XhoI-ApaI-KpnI"
FT   misc_binding    0..0
FT                   /note="SIT unique SacI-SacII-EagI-NotI-SpeI-BamHI-
FT                   SmaI-PstI-HindIII-SalI-XhoI-ApaI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T3"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T7"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
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"
FT   rep_origin      0..0
FT                   /note="ORI yeast 2 micron"
XX
SQ   Sequence 6849 BP; 1869 A; 1504 C; 1543 G; 1933 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
     accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
     ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
     taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
     ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
     aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
     agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
     tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
     ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
     ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
     tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
     aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
     ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
     aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     acagtttttc tccataatct tgaagaggcc aaaagattag ctttatccaa ggaccaaata
     ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     ctaccggtac cccatttagg accagccaca gcacctaaca aaacggcatc aaccttcttg
     gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc
     ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt
     ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg
     taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc
     cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 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 gccacctgaa cgaagcatct gtgcttcatt
     ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
     atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
     gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
     cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
     cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
     gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
     catactaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     cctttcgtc
//