ID PRS316 preliminary; circular DNA; SYN; 4887 BP.
XX
AC U03442; ATCC77145;
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 pRS316 - complete.
XX
KW cloning vector.
XX
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).
XX
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).
XX
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.
XX
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"
XX
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
//