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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"
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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
//