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ID PRS313 preliminary; circular DNA; SYN; 4967 BP.
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AC U03439; ATCC77142;
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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 pRS313 - 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 1-4967
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).
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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.
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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 ()
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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/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
//