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ID   YIPLAC211  preliminary; circular DNA; SYN; 3797 BP.
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AC   X75462; L26358;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector YIplac211 - 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   1-3797
RC   YIplac211
RA   Gietz R.D.;
RT   ;
RL   Submitted (13-MAY-1993) by:
RL   Gietz R.D., Health Science Ctr., University of Manitoba,
RL   Manitoba, CANADA.
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RN   [2]
RP   1-3797
RC   plasmid1 from pIBI30 & M13mp19 & YEp24, URA3 gene
RC   plasmid2 from pIBI30 & M13mp19 & YRp7, TRP1/ARS1 gene
RC   plasmid3 from pIBI30 & M13mp19 & YEp213, LEU2 gene
RC   YCplac22 from pUC19 & pTC1
RC   pAC5 from pBR322 & pTC1
RC   YCplac33 from plasmid1 & pAC5
RC   YCplac111 from plasmid3 & pAC5
RC   p2micron19 from pBR322 & YEp24
RC   YEplac112 from p2micron19 & plasmid1
RC   YEplac181 from p2micron19 & plasmid2
RC   YEplac195 from p2micron19 & plasmid3
RC   YIplac128 from pUC19 & plasmid1
RC   YIplac204 from pUC19 & plasmid2
RC   YIplac211 from pUC19 & plasmid3
RA   Gietz R.D., Sugino A.;
RT   "New yeast-Escherichia coli shuttle vectors constructed with in
RT   vitro mutagenized yeast genes lacking six-base pair restriction
RT   sites";
RL   Gene 74:527-534(1988).
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CC   NOTE: Information taken from figure 2, citation [2].
CC   In E. coli plasmid pBR322, insert yeast genes into either EcoRI or
CC   Cla I using PolIK and dNTPs to fill in overhangs and T4 DNA ligase
CC   for blunt-end ligation.
CC   Any restriction site regenerated by this process was destroyed
CC   following digestion with the appropriate restriction enzyme,
CC   using PolIK and dNTPs to fill in overhangs and T4 DNA ligase to
CC   ligate blunt ends.
CC   Once the constructions were verified, the yeast genes were moved into
CC   the plasmid vector pUC19.
CC   Excise the gene from pBR322 by HindIII digestion, followed by
CC   mung-bean exonuclease treatment to produce blunt ends.
CC   Construct YCplac vectors (ARS1-CEN4) by blunt-end ligation of the
CC   1.4-kb EcoRI fragment (TRP1 & ARS1 genes) [3] into pBR322 EcoRI
CC   site (to make pT322) and subsequent blunt-end ligation of the
CC   functional 850-bp PvuII-HpaI CEN4 fragment [4] into pT322 ClaI
CC   (to make pTC1).
CC   Digest with AatII, and isolate the fragment containing yeast DNA
CC   from agarose gels and ligate to the large AatII-Nde I fragment of
CC   pUC19, which had the NdeI end filled in with PolIK (to make YCplac22).
CC   Make the other YCplac vectors by blunt-end ligation of the EcoRV
CC   fragment from pTC1, containing the ARS1 and CEN4 sequences, into ClaI
CC   pBR322 ClaI (to make pAC5).
CC   Insert the 1.1-kb HindIII-SmaI fragment (containing URA3)
CC   or the 1.6-kb HpaI-AccI fragment (containing LEU2) at pAC5 EcoRI
CC   as described above.
CC   Move these constructs into pUC19 as described above, to make
CC   YCplac33 and YCplac111, respectively.
CC   Construct the YEplac vector series (with a yeast 2 mu origin)
CC   by blunt-end ligation of the 1.5-kb SauIII fragment
CC   from YEp24, which contains the yeast 2 mu origin [5], into pBR322
CC   ClaI (to make p2mu19).
CC   Remove p2mu19 XbaI site by filling in with PolIK after digestion and
CC   blunt-end religation of this site.
CC   Ligate the three yeast genes into p2mu19 EcoRI. The fragments used
CC   for the LEU2 and the URA3 constructs were the same as those used in
CC   the YCplac constructions. However, the YEplac112 vector (TRP1)
CC   contains the 850-bp EcoRI-BglII fragment of the TRP1 gene which does
CC   not contain ARS1 function.
CC   Construct the YIplac vectors by ligating each yeast gene fragment
CC   used in YEplac vector constructions into the pUC19 EcoO109 site
CC   using PolIK to fill the sticky ends.
CC   NCBI gi: 415335
CC   NM (YIplac211)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)(Saccharomyces cerevisiae)
CC   CP ()
CC   FN (cloning)(shuttle)
CC   SE ()
CC   PA (pUC19)(pBR322)(yeast 2-micron plasmid)
CC   BR (YCplac22)(YCplac33)(YCplac111)(YEplac112)(YEplac181)(YEplac195)
CC   BR (YIplac128)(YIplac204)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. M13mp19 remove HindIII-SmaI 37bp 6235..6272,
FT                   \ 7213bp
FT                   2. YEp24 HindIII-SmaI 1108bp 2273..3381, URA3 gene
FT                   -> plasmid 8321bp
FT                   1. plasmid PstI 8321bp, YEp24 3544..3544
FT                   oligo mutagenesis
FT                   -> plasmid2 8321bp
FT                   1. pIBI30 remove SmaI-HindIII 51bp 286..337, 2875bp
FT                   2. plasmid2 HindIII-SmaI 1108bp 2273..3381, URA3 gene
FT                   -> plasmid3 3983bp
FT                   1. pUC19 EcoO109 2686bp 2676..2676
FT                   2. plasmid3 HindIII-SmaI 1108bp 2273..3381, URA3 gene
FT                   -> YIplac211 3797bp"
FT   misc_feature    0..0
FT                   /note="from pBR322"
FT   misc_feature    0..0
FT                   /note="from pUC19"
FT   misc_feature    0..0
FT                   /note="poly dG-dC tail"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
FT   misc_feature    0..0
FT                   /note="poly dG-dC tail"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN4"
FT   misc_binding    0..0
FT                   /note="MCS from pUC19"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARS1"
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 E. coli tetracycline resistance gene (tet)"
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SQ   Sequence 3797 BP; 973 A; 945 C; 858 G; 1021 T; 0 other;
     gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
     atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
     ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     caccgtcatc accgaaacgc gcgagacgaa agggcgggta ataactgata taattaaatt
     gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct gtgcagttgg
     gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa
     gctggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     agcgaggaag cggaaga
//