LOCUS       40924_49617        8434 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal vector with a LEU2 marker for construction of lacZ 
            fusions. The MCS is reversed in YEp366.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8434)
  AUTHORS   Myers AM, Tzagoloff A, Kinney DM, Lusty CJ.
  TITLE     Yeast shuttle and integrative vectors with multiple cloning sites 
            suitable for construction of lacZ fusions.
  JOURNAL   Gene 1986;45:299-310.
  PUBMED    3026915
REFERENCE   2  (bases 1 to 8434)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8434)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1986"; volume: "45"; pages: "299-310"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..57)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             73..3120
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLR
                     SLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNP
                     PFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSE
                     FDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVAT
                     RFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADR
                     VTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGK
                     PLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYG
                     LYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHG
                     ANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWL
                     SLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGN
                     PWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEY
                     LFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVV
                     QPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNR
                     QSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAE
                     AALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPH
                     PARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGL
                     RCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGD
                     DSWSPSVSAELQLSAGRYHYQLVWCQK"
     CDS             complement(3716..4807)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MFVPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWTLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEALRYPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPGFAKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4808..5212)
                     /label=LEU2 promoter
     rep_origin      complement(5213..6195)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6477..6581
                     /label=AmpR promoter
     CDS             6582..7439
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7613..8201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattccca
       61 gcttgcgatc ccgccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      121 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      181 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
      241 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
      301 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgta
      361 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
      421 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
      481 gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag
      541 gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg agaaaaccgc
      601 ctcgcggtga tggtgctgcg ttggagtgac ggcagttatc tggaagatca ggatatgtgg
      661 cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac acaaatcagc
      721 gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact ggaggctgaa
      781 gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt atggcagggt
      841 gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga tgagcgtggt
      901 ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact gtggagcgcc
      961 gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt
     1021 gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg
     1081 ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca tcctctgcat
     1141 ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa gcagaacaac
     1201 tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac gctgtgcgac
     1261 cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat ggtgccaatg
     1321 aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg
     1381 gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa tgaatcaggc
     1441 cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg
     1501 gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg cccgatgtac
     1561 gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat caaaaaatgg
     1621 ctttcgctac ctggagagac gcgcccgctg atcctttgcg aatacgccca cgcgatgggt
     1681 aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc ccgtttacag
     1741 ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga aaacggcaac
     1801 ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca gttctgtatg
     1861 aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc aaaacaccag
     1921 cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag cgaatacctg
     1981 ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg taagccgctg
     2041 gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat tgaactgcct
     2101 gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg
     2161 aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg gcgtctggcg
     2221 gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct gaccaccagc
     2281 gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg ccagtcaggc
     2341 tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct gcgcgatcag
     2401 ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg cattgaccct
     2461 aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc agcgttgttg
     2521 cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag
     2581 catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg tagtggtcaa
     2641 atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc gcggattggc
     2701 ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt agggccgcaa
     2761 gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct gccattgtca
     2821 gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa
     2881 ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag ccgctacagt
     2941 caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga aggcacatgg
     3001 ctgaatatcg acggtttcca tatggggatt ggtggcgacg actcctggag cccgtcagta
     3061 tcggcggaat tacagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa
     3121 taataataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc cattatgtac
     3181 tatttcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     3241 atccggtacg tcgttaaggc cgtttctgac agagtaaaat tcttgaggga actttcacca
     3301 ttatggaaaa tggttcaaga aggtattgac ttaaactcca tcaaatggtc aggtcatttg
     3361 agtgtttttt atttgttgta tttttttttt tttagagaaa atcctccaat atcaaattag
     3421 ggattcgtag tttcatgatt ttctgttata cctaactttt tgtgtggtgc cctcctcctt
     3481 gtcaatatta atgttaaagt gcaattctct ttccttatca cgttgagcca ttagtatcaa
     3541 tttgcttacc tgtattcctt tagctatcct cctttttctc cttcttgata aatgtatgta
     3601 gattgcgttt gtctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
     3661 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aagagaatct ttttaagcaa
     3721 ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg ttggaaccac
     3781 ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct tcaatggcct
     3841 taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac aagatagtgg
     3901 cgatagggtc aaccttattc ttggcaaatc cgggagcaga accgtggcat ggttcgtaca
     3961 aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc aacaaaccca
     4021 aggaacctgg ataacgtaag gcttcatcgg agatgatatc accaaacatg ttgctggtga
     4081 ttataatacc atttaggtgg gttgggttct taactaggat catggcggca gaatcaatca
     4141 attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc acagtttttc
     4201 tccataatct tgaagaggcc aaaacattag ctttatccaa ggtccaaata ggcaatggtg
     4261 gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact tctggaacgg
     4321 tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc ttaccaaagt
     4381 aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca aattgtggct
     4441 tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt aagttggcgt
     4501 acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca ctaccggtac
     4561 cccattttgg accacccaca gcacctaaca aaacggcatc aaccttcttg gaggcttcca
     4621 gtgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca attaaatgat
     4681 tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga accttaatgg
     4741 cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc ttcttaggaa
     4801 caaacattag aatggtatat ccttgaaata tatatatata ttgctgaaat gtaaaaggta
     4861 agaaaagtta gaaagtaaga cgattgctaa ccacctattg gaaaaaacaa taggtcctta
     4921 aataatattg tcaacttcaa gtattgtgat gcaagcattt agtcatgaac gcttctctat
     4981 tctatatgaa aagccggttc cggcctctca cctttccttt ttctcccaat ttttcagttg
     5041 aaaaaggtat atgcgtcagg cgacctctga aattaacaaa aaatttccag tcatcgaatt
     5101 tgattctgtg cgatagcgcc cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt
     5161 gctaagagat tcgaactctt gcatcttacg atacctgagt attcccacag ttaacgaagc
     5221 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     5281 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     5341 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     5401 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     5461 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     5521 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     5581 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     5641 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     5701 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     5761 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     5821 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     5881 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     5941 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     6001 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     6061 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     6121 ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt
     6181 tcctatactt tctagtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6241 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6301 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6361 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     6421 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     6481 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6541 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6601 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6661 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6721 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6781 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6841 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6901 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6961 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     7021 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     7081 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     7141 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7201 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7261 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7321 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7381 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7441 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7501 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7561 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7621 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7681 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7741 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7801 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7861 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7921 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7981 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     8041 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8101 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8161 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8221 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8281 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8341 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     8401 cgcctctccc cgcgcgttgg ccgattcatt cccg
//
