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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.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
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).
XX
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).
XX
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.
XX
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
//