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ID   PRS316     preliminary; circular DNA; SYN; 4887 BP.
XX
AC   U03442; ATCC77145;
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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 phagemid vector pRS316 - 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   1-4887
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   from YEp24
RC   from YCp50
RC   from YIp5
RA   Strathern J.N., Higgins D.R.;
RT   "Recovery of plasmids from yeast into Escherichia coli: shuttle
RT   vectors";
RL   Meth. Enzymol. 194:319-329(1991).
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RN   [3]
RC   pRS316
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. [1]
CC   The order of the major features in this plasmid is: URA3 - f1 ori
CC   (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla -
CC   CEN6 - ARSH4.
CC   A fragment (1.112 kb) containing the URA3 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(+). [1]
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. [1]
CC   Useful in plasmid shuffle experiments.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--5.0; BamHI--5.0, PvuII--4.4, 0.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 417973
CC   NM (pRS316)
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)(pRS315)(pRS313)
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. pRS306 AatII 4373bp 848..848
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pRS316 4887bp"
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 KpnI-ApaI-XhoI-SalI-ClaI-EcoRI-
FT                   PstI-SmaI-BamHI-SpeI-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 URA3 gene"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARSH4"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
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SQ   Sequence 4887 BP; 1361 A; 1069 C; 1167 G; 1290 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
     tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
     ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg
     ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc
     cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
     gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
     aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
     actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga
     gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat 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 gactgcccgt 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
     gtgaaaaaaa 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 cctgggtcct tttcatcacg
     tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
     ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa
     ataggcgtat cacgaggccc tttcgtc
//