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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.
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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 ()
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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. 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
//