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ID   YIPLAC204  preliminary; circular DNA; SYN; 3545 BP.
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AC   X75461; L26357;
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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 YIplac204 - 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   1-3545
RC   YIplac204
RA   Gietz R.D.;
RT   ;
RL   Submitted (13-MAY-1993) by:
RL   Gietz R.D., Health Sciuence Ctr., University of Manitoba,
RL   Manitoba, CANADA.
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RN   [2]
RP   1-3545
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: 415334
CC   NM (YIplac204)
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)(YIplac211)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pUC19 EcoO109 2686bp 2676..2676
FT                   Klenow:Klenow
FT                   2. YRp7 BglII-EcoRI 852bp 604..1456, TRP1
FT                   Klenow:Klenow
FT                   -> YIplac204 3545bp"
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 TRP1 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 3545 BP; 899 A; 906 C; 817 G; 923 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 agggcgatct tttatgcttg cttttcaaaa
     ggcttgcagg caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt
     cttagcattt ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac
     acctccgctt acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc
     tggcgtcagt ccaccagcta acataaaatg taagctctcg gggctctctt gccttccaac
     ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa
     caagggaata aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt
     tggaaatacg agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga
     ctcatctcca tgcagttgga cgatatcaat gccgtaatca ttgaccagag ccaaaacatc
     ctccttaggt tgattacgaa acacgccaac caagtatttc ggagtgcctg aactattttt
     atatgctttt acaagacttg aaattttcct tgcaataacc gggtcaattg ttctctttct
     attgggcaca catataatac ccagcaagtc agcatcggaa tctagtgcac attctgcggc
     ctctgtgctc tgcaagccgc aaactttcac caatggacca gaactacctg tgaaattaat
     aacagacata ctccaagctg cctttgtgtg cttaatcacg tatactcacg tgctcaatag
     tcaccaatgc cctccctctt ggccctctcc ttttcttttt tcgaccgaat tggcctcgtg
     atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     gaaga
//