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ID   YIPLAC128  preliminary; circular DNA; SYN; 4302 BP.
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AC   X75463; L26356;
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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 YIplac128 - 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   1-4302
RC   YIplac128
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   1-4302
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: 415333
CC   NM (YIplac128)
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 (YIplac204)(YIplac211)
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. M13mp19 remove BglI-BglII 498bp 6438..6936,
FT                   \ 6752bp
FT                   2. YEp213 BglI-BglII 1442bp 3492..4934, LEU2 gene
FT                   -> plasmid 8194bp
FT                   1. plasmid KpnI 8194bp, M13mp19 6278..6278
FT                   oligo mutagenesis
FT                   EcoRI 8194bp, M13mp19 6286..6286
FT                   oligo mutagenesis
FT                   -> plasmid2 8194bp
FT                   1. pIBI30 AccI 2926bp 302..302
FT                   2. plasmid2 HpaI-AccI 1600bp, LEU2 gene
FT                   -> plasmid3 4500bp
FT                   1. plasmid3 HpaI-AccI 1600bp, LEU2 gene
FT                   blunt end:blunt end
FT                   2. pUC19 EcoO109 2686bp 2676..2676
FT                   blunt end:blunt end
FT                   -> YIplac128 4302bp"
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 LEU2 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 4302 BP; 1141 A; 1027 C; 993 G; 1141 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 agggcctacc ctatgaacat attccatttt
     gtaatttcgt gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata
     aaaaaagaga atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga
     cttcggtggt actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct
     ttttaactgc atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca
     tcattgcagc agacaagata gtggcgatag ggtcaacctt attctttggc aaatctggag
     cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca
     aggacgcaga tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga
     tatcaccaaa catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta
     ggatcatggc ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct
     tgatggtttc ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat
     ccaaggacca aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg
     tgattctttg cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat
     cgtcttcctt tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt
     cagtaccttt agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa
     agttacatgg tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt
     gttcaggtct aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg
     catcaacctt cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga
     tagcagcacc accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag
     aaatagcttt aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg
     gcaaaacgac gatcttctta ggggcagaca taggggcaga cattagaatg gtatatcctt
     gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat
     tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat
     tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc
     ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc gtcaggcgac
     ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat agcgcccctg
     tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga actcttgcat
     cttacgatac ctgagtattc ccacagttgg cctcgtgata cgcctatttt tataggttaa
     tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa ga
//