back Return to this vector's summary.
ID   PRS313     preliminary; circular DNA; SYN; 4967 BP.
XX
AC   U03439; ATCC77142;
XX
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   Saccharomyces/E.coli phagemid vector pRS313 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-4967
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   [2]
RC   pRS313
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   One of a series of pBluescript-based centromere vectors (ATCC 77142
CC   to 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: HIS3 - f1 ori
CC   (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla -
CC   CEN6 - ARSH4.
CC   A fragment (1.184 kb) containing the HIS3 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--5.0; BamHI--5.0; PvuII--4.5, 0.48. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416308
CC   NM (pRS313)
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)(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/MboI-Sau3AI/MboI 374bp,
FT                   \ 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. pRS303 AatII 4443bp 4389..4389
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pRS313 4967bp"
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-ClaI-EcoRV-EcoRI-
FT                   SmaI-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 HIS3 gene"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARSH4"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
XX
SQ   Sequence 4967 BP; 1335 A; 1132 C; 1171 G; 1329 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accataattc cgttttaaga gcttggtgag cgctaggagt cactgccagg tatcgtttga
     acacggcatt agtcagggaa gtcataacac agtcctttcc cgcaattttc tttttctatt
     actcttggcc tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt
     ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
     tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
     ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc gtattacaaa
     tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga tagagcactc
     gatcttccca gaaaaagagg cagaagcagt agcagaacag gccacacaat cgcaagtgat
     taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg ccaagcattc
     cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc atcacaccac
     tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctactgg cgcgtggagt
     aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggtaga
     tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggaga
     tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac
     cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa
     ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc
     caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat acgatatata
     tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa cagtatgata
     ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag gcgcgctttc
     cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatca tatgcggtgt
     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
//