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ID   PRS314     preliminary; circular DNA; SYN; 4783 BP.
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AC   U03440; ATCC77143;
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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 pRS314 - 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   1-4783
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   pRS314
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: TRP1 - f1 ori
CC   (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla -
CC   CEN6 - ARSH4.
CC   A genomic HincII/PstI fragment (1.002 kb) containing the TRP1 gene was
CC   inserted into the NdeI site and a cassette containing CEN6 and the ARS
CC   associated with histone 4 (ARSH4) was inserted into the AatII site of
CC   pRSS56.  All ends were blunted.
CC   An EcoRI site in the TRP1-containing fragment (external to the coding
CC   sequence) was destroyed.
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--4.8; BamHI--4.8; PvuII--4.2, 0.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416309
CC   NM (pRS314)
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 (pRS313)(pRS315)(pRS316)
CC   OF ()
CC   OR ()
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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. pRS304 AatII 4267bp 4203..4203
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pRS314 4783bp"
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 TRP1 gene"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARSH4"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
XX
SQ   Sequence 4783 BP; 1309 A; 1047 C; 1153 G; 1274 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
     atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
     aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
     gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
     tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
     atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
     cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
     agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
     atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
     ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
     ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
     actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
     attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
     gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgcctg cggtgtgaaa
     taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt
     gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg caaggcgatt
     aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggcggcc gctctagaac
     tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga
     gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattccg agcttggcgt
     aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
     taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     tcggcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     caggactata aagataccag gcgttccccc ctggaagctc cctcgtgcgc tctcctgttc
     cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     cagcgatctg tctatttcgt tcatccatag ttgcctgact gcccgtcgtg tagataacta
     cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     catgatcccc catgttgtga aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata
     taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa
     gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc
     tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat
     cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc
     ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt
     attttttttt cttcattccg taactcttct accttcttta tttactttct aaaatccaaa
     tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag
     atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct aagaaaccat tattatcatg
     acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//