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ID   PRS315     preliminary; circular DNA; SYN; 6018 BP.
XX
AC   U03441; ATCC77144;
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 pRS315 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-6018
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).
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RN   [2]
RC   pRS315
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.
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CC   One of a series of pBluescript-based centromere vectors (ATCC 77142
CC   ATCC 77145, ATCC 77157 to ATCC 77158) differing in the yeast
CC   selectable marker gene.
CC   The order of the major features in this plasmid is: LEU2 - f1 ori
CC   (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla -
CC   CEN6 - ARSH4.
CC   A fragment (2.235 kb) containing the LEU2 gene was inserted into the
CC   NdeI site and a cassette containing CEN6 and the ARS associated with
CC   histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends
CC   were blunted.
CC   pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of
CC   pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains
CC   the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites
CC   between bla and f1 origin of pBS(+).
CC   YC-type centromere vector permitting visual detection of recombinants
CC   and production of ssDNA in E.coli.  Contains promoters for in vitro
CC   RNA synthesis, priming sites useful for sequencing, and encodes the
CC   lacZ alpha (lacZ') peptide.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--3.9,2.1; BamHI--6.0; HindIII--6.0; KpnI--4.3, 1.7;
CC   KpnI/EcoRI--4.1, 1.5, 0.4. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416310
CC   NM (pRS315)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
CC   HO (Saccharomyces cerevisiae YPH499)(Saccharomyces cerevisiae YPH500)
CC   HO (Saccharomyces cerevisiae YPH501)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR (pRS314)(pRS313)(pRS316)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC19 HincII 2686bp 432..432
FT                   2. yeast MboII-MboII 125bp, CEN6
FT                   -> pUC19-CEN6:32 2811bp
FT                   1. pUC8 BamHI 2665bp 241..241
FT                   2. yeast Sau3AI-Sau3AI 374bp, histone H4 ARS/ARSH4
FT                   -> pAB9 3039bp
FT                   1. pUC19-CEN6:32 EcoRI-ScaI 1908bp 397..-..2180, CEN6
FT                   2. pAB9 ScaI-EcoRI 1314bp 231..-..1171,
FT                   \ histone H4 ARS/ARSH4
FT                   -> pRSS83 3222bp [in MCS]
FT                   1. pRSS83 remove SmaI-XbaI 9bp, MCS/pUC19 415..424
FT                   \ 3213bp
FT                   Klenow:Klenow
FT                   -> pRSS84 3213bp
FT                   1. pRSS84 HincII-PstI, CEN6/ARSH4
FT                   \ pUC19 440..2686..432
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   2. pRS305 AatII 5504bp 5440..5440
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pRS315 6018bp"
FT   misc_binding    0..0
FT                   /note="MCS KpnI-ApaI-XhoI-SalI-ClaI-HindIII-EcoRV-
FT                   EcoRI-PstI-SmaI-
FT                   BamHI-SpeI-XbaI-EagI-NotI-BstXI-SacII-SacI"
FT   misc_binding    0..0
FT                   /note="SIT unique ApaI-XhoI-SalI-HindIII-PstI-SmaI-
FT                   BamHI-SpeI-XbaI-EagI-NotI-SacII-SacI-NaeI"
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 ARSH4"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
XX
SQ   Sequence 6018 BP; 1666 A; 1328 C; 1341 G; 1683 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 gaaatggttc 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 aaaacattag 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 accacccaca 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 tcgagggtcg 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 aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa
     ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg
     ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt
     gtgaaattgt tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa
     agcctggggt gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc
     tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     taaaaaggcc gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa
     aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     cccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     catagttgcc tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg
     ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa
     agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
     aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
     agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
     ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
     tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
     gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
     tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
     cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
     cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
     caagcgatcc gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     tcacgaggcc ctttcgtc
//