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ID   PRS424     preliminary; circular DNA; SYN; 5616 BP.
XX
AC   U03453; ATCC77105;
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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 pRS424 - 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   1-5616
RC   plasmid from YEp24 & pRSS56
RC   [pBR322/SC4128 from pBR322 & yeast]
RC   pJS92 from plasmid & pBR322/SC4128
RC   pRS323 from pJS92 & pRS303
RC   pRS324 from pJS92 & pRS304
RC   pRS325 from pJS92 & pRS305
RC   pRS326 from pJS92 & pRS306
RC   pRS423 from pRS323 & pBluescript II SK+
RC   pRS424 from pRS324 & pBluescript II SK+
RC   pRS425 from pRS325 & pBluescript II SK+
RC   pRS426 from pRS326 & pBluescript II SK+
RA   Christianson T.W., Sikorski R.S., Dante M., Shero J.H., Hieter P.;
RT   "Multifunctional yeast high-copy-number shuttle vectors";
RL   Gene 110:119-122(1992).
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RN   [2]
RP   1-5616
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   [3]
RC   pRS424
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   The order of the major features in this plasmid is:ori(f1) - lacZ - T7
CC   promoter - MCS (KpnI-SacI) - T3 promoter - lacI - ori(pMB1) - ampR -
CC   ori (2 micron) - TRP1.
CC   Contains the REP3 and FRT sequences necessary for high copy
CC   propagation in yeast.
CC   In S. cerevisiae, the number is about 20 per haploid cell.  In
CC   non-selective growth, plasmids are lost through mitotic segregation at
CC   rates in the range of 1.5 + or - 1.7% of progeny per doubling.
CC   YE-type shuttle vector encoding the beta-galactosidase alpha peptide,
CC   permitting blue-white visual detection. Can be used to produce ssDNA.
CC   Contains promoters for in vitro RNA synthesis and priming sites useful
CC   for sequencing.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.6; EcoRI--5.6; PvuII--5.2, 0.5; XbaI--3.3, 1.4, 1.0. (ATCC
CC   staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416324
CC   NM (pRS424)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli DH5alpha)(Saccharomyces cerevisiae)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR (pRS423)(pRS425)(pRS426)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. YEp24 HpaI-NdeI 1346bp 341..1687,
FT                   \ yeast 2 micron ori [1345bp]
FT                   :Klenow
FT                   2. pRSS56 AatII 2998bp 3140..3140
FT                   T4 DNA polymerase
FT                   -> plasmid 4344bp [unique NdeI]
FT                   1. pBR322 4361bp
FT                   2. yeast DNA 1800bp, TRP1 gene
FT                   -> pBR322/SC4128 6200bp
FT                   1. plasmid NdeI 4344bp 185..185
FT                   Klenow
FT                   2. pBR322/SC4128 BglII-BglII 1800bp, TRP1 gene
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   -> pJS92 6250bp
FT                   1. pJS92 remove DraII-DraII 1800bp, TRP1 gene/4445bp
FT                   2. pRS304 DraII-DraII 2273bp 1984..4257, TRP1 gene
FT                   -> pRS324 5613bp
FT                   1. pRS324 remove PvuI-PvuI 1913bp 1738..3651,
FT                   \ MCS/3700bp
FT                   2. pBluescript II SK+ PvuI-PvuI 1916bp 501..2417, MCS
FT                   -> pRS424 5616bp"
FT   misc_binding    0..0
FT                   /note="MCS SacI-BstXI-SacII-EagI-NotI-XbaI-SpeI-
FT                   BamHI-SmaI-PstI-
FT                   EcoRI-EcoRV-HindIII-ClaI-SalI-XhoI-ApaI-KpnI"
FT   misc_binding    0..0
FT                   /note="SIT unique SacI-SacII-EagI-NotI-SpeI-BamHI-
FT                   SmaI-PstI-EcoRI-ClaI-SalI-XhoI-ApaI-KpnI"
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 2 micron"
XX
SQ   Sequence 5616 BP; 1513 A; 1221 C; 1356 G; 1526 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 cacttgccta tgcggtgtga
     aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
     ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg
     gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     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
     taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     gctttctcat agctcacgct 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 tgactccccg 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 tgtgcaaaaa 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
     acctgaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag
     cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     atgctatcat ttcctttgat attggatcat attaagaaac cattattatc atgacattaa
     cctataaaaa taggcgtatc acgaggccct ttcgtc
//