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ID   PRS423     preliminary; circular DNA; SYN; 5797 BP.
XX
AC   U03454; ATCC77104;
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 pRS423 - complete.
XX
KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-5797
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   1-5797
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   pRS423
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   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   In S. cerevisiae, the copy 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 4.3 + or - 1.3% of progeny per doubling.
CC   Contains the REP3 and FRT sequences necessary for high copy
CC   propagation in yeast.
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) - HIS3.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--6.0; EcoRI--6.0; PvuII--5.4, 0.52; XbaI--3.3, 2.7. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NCBI gi: 416325
CC   NM (pRS423)
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 (pRS424)(pRS425)(pRS426)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. YEp24 NdeI-HpaI 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 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. pRS303 DraII-DraII 1349bp 821..2170, HIS3 gene
FT                   -> pRS323 5794bp
FT                   1. pRS323 remove PvuI-PvuI 1913bp 1924..3837,
FT                   \ MCS/3881bp
FT                   2. pBluescript II SK+ PvuI-PvuI 1916bp 501..2417, MCS
FT                   -> pRS423 5797bp"
FT   misc_binding    0..0
FT                   /note="MCS SacI-BstXI-SacII-EagI-NotI-XbaI-SpeI-
FT                   BamHI-SmaI-PstI-EcoRI-EcoRV-HindIII-ClaI-SalI-XhoI-
FT                   ApaI-KpnI"
FT   misc_binding    0..0
FT                   /note="SIT unique SacI-SacII-EagI-NotI-SpeI-BamHI-
FT                   SmaI-EcoRI-EcoRV-ClaI-SalI-XhoI-ApaI"
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 2 micron"
XX
SQ   Sequence 5797 BP; 1536 A; 1308 C; 1374 G; 1579 T; 0 other;
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
     gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
     ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
     tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
     atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
     aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
     atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
     cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
     ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
     ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
     ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
     taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
     atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
     atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc 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 agggggatgt gctgcaaggc
     gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact agttctagag
     cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     cacaacatag gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa
     ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     ccacctgaac gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct
     aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc
     gctattttac caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga
     gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     agagcgctat tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc
     cgagagcgct atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc
     tataatgcag tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct
     actttggtgt ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg
     attactagcg aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta
     taccgatgtg gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat
     tggtcagaaa attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat
     gtttacattt tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt
     gtctaaagag taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag
     ttcaaggagc gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc
     aaagagatac ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt
     tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt ttggttttca
     aaagcgctct gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa
     agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac
     tgttcacgtc gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg
     catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag
     gtagtctagt acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc
     agcactaccc tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt
     caatgctatc atttcctttg atattggatc atctaagaaa ccattattat catgacatta
     acctataaaa ataggcgtat cacgaggccc tttcgtc
//