LOCUS       40924_49632        8426 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 YEp368R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8426)
  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 8426)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8426)
  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..8426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             65..3112
                     /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(3708..4799)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MFVPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWTLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEALRYPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPGFAKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4800..5204)
                     /label=LEU2 promoter
     rep_origin      complement(5205..6187)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6469..6573
                     /label=AmpR promoter
     CDS             6574..7431
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7605..8193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttcga
       61 tcccgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      121 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      181 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
      241 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
      301 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg taacctatcc
      361 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
      421 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
      481 taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc ggttacggcc aggacagtcg
      541 tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc ggagaaaacc gcctcgcggt
      601 gatggtgctg cgttggagtg acggcagtta tctggaagat caggatatgt ggcggatgag
      661 cggcattttc cgtgacgtct cgttgctgca taaaccgact acacaaatca gcgatttcca
      721 tgttgccact cgctttaatg atgatttcag ccgcgctgta ctggaggctg aagttcagat
      781 gtgcggcgag ttgcgtgact acctacgggt aacagtttct ttatggcagg gtgaaacgca
      841 ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc gatgagcgtg gtggttatgc
      901 cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa ctgtggagcg ccgaaatccc
      961 gaatctctat cgtgcggtgg ttgaactgca caccgccgac ggcacgctga ttgaagcaga
     1021 agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat ggtctgctgc tgctgaacgg
     1081 caagccgttg ctgattcgag gcgttaaccg tcacgagcat catcctctgc atggtcaggt
     1141 catggatgag cagacgatgg tgcaggatat cctgctgatg aagcagaaca actttaacgc
     1201 cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac acgctgtgcg accgctacgg
     1261 cctgtatgtg gtggatgaag ccaatattga aacccacggc atggtgccaa tgaatcgtct
     1321 gaccgatgat ccgcgctggc taccggcgat gagcgaacgc gtaacgcgaa tggtgcagcg
     1381 cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg aatgaatcag gccacggcgc
     1441 taatcacgac gcgctgtatc gctggatcaa atctgtcgat ccttcccgcc cggtgcagta
     1501 tgaaggcggc ggagccgaca ccacggccac cgatattatt tgcccgatgt acgcgcgcgt
     1561 ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc atcaaaaaat ggctttcgct
     1621 acctggagag acgcgcccgc tgatcctttg cgaatacgcc cacgcgatgg gtaacagtct
     1681 tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac agggcggctt
     1741 cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca acccgtggtc
     1801 ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta tgaacggtct
     1861 ggtctttgcc gaccgcacgc cgcatccagc gctgacggaa gcaaaacacc agcagcagtt
     1921 tttccagttc cgtttatccg ggcaaaccat cgaagtgacc agcgaatacc tgttccgtca
     1981 tagcgataac gagctcctgc actggatggt ggcgctggat ggtaagccgc tggcaagcgg
     2041 tgaagtgcct ctggatgtcg ctccacaagg taaacagttg attgaactgc ctgaactacc
     2101 gcagccggag agcgccgggc aactctggct cacagtacgc gtagtgcaac cgaacgcgac
     2161 cgcatggtca gaagccgggc acatcagcgc ctggcagcag tggcgtctgg cggaaaacct
     2221 cagtgtgacg ctccccgccg cgtcccacgc catcccgcat ctgaccacca gcgaaatgga
     2281 tttttgcatc gagctgggta ataagcgttg gcaatttaac cgccagtcag gctttctttc
     2341 acagatgtgg attggcgata aaaaacaact gctgacgccg ctgcgcgatc agttcacccg
     2401 tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc cgcattgacc ctaacgcctg
     2461 ggtcgaacgc tggaaggcgg cgggccatta ccaggccgaa gcagcgttgt tgcagtgcac
     2521 ggcagataca cttgctgatg cggtgctgat tacgaccgct cacgcgtggc agcatcaggg
     2581 gaaaacctta tttatcagcc ggaaaaccta ccggattgat ggtagtggtc aaatggcgat
     2641 taccgttgat gttgaagtgg cgagcgatac accgcatccg gcgcggattg gcctgaactg
     2701 ccagctggcg caggtagcag agcgggtaaa ctggctcgga ttagggccgc aagaaaacta
     2761 tcccgaccgc cttactgccg cctgttttga ccgctgggat ctgccattgt cagacatgta
     2821 taccccgtac gtcttcccga gcgaaaacgg tctgcgctgc gggacgcgcg aattgaatta
     2881 tggcccacac cagtggcgcg gcgacttcca gttcaacatc agccgctaca gtcaacagca
     2941 actgatggaa accagccatc gccatctgct gcacgcggaa gaaggcacat ggctgaatat
     3001 cgacggtttc catatgggga ttggtggcga cgactcctgg agcccgtcag tatcggcgga
     3061 attacagctg agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataataata
     3121 accgggcagg ccatgtctgc ccgtatttcg cgtaaggaaa tccattatgt actatttcgc
     3181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agatccggta
     3241 cgtcgttaag gccgtttctg acagagtaaa attcttgagg gaactttcac cattatggaa
     3301 aatggttcaa gaaggtattg acttaaactc catcaaatgg tcaggtcatt tgagtgtttt
     3361 ttatttgttg tatttttttt tttttagaga aaatcctcca atatcaaatt agggattcgt
     3421 agtttcatga ttttctgtta tacctaactt tttgtgtggt gccctcctcc ttgtcaatat
     3481 taatgttaaa gtgcaattct ctttccttat cacgttgagc cattagtatc aatttgctta
     3541 cctgtattcc tttagctatc ctcctttttc tccttcttga taaatgtatg tagattgcgt
     3601 ttgtctaccc tatgaacata ttccattttg taatttcgtg tcgtttctat tatgaatttc
     3661 atttataaag tttatgtaca aatatcataa aaaagagaat ctttttaagc aaggattttc
     3721 ttaacttctt cggcgacagc atcaccgact tcggtggtac tgttggaacc acctaaatca
     3781 ccagttctga tacctgcatc caaaaccttt ttaactgcat cttcaatggc cttaccttct
     3841 tcaggcaagt tcaatgacaa tttcaacatc attgcagcag acaagatagt ggcgataggg
     3901 tcaaccttat tcttggcaaa tccgggagca gaaccgtggc atggttcgta caaaccaaat
     3961 gcggtgttct tgtctggcaa agaggccaag gacgcagatg gcaacaaacc caaggaacct
     4021 ggataacgta aggcttcatc ggagatgata tcaccaaaca tgttgctggt gattataata
     4081 ccatttaggt gggttgggtt cttaactagg atcatggcgg cagaatcaat caattgatgt
     4141 tgaaccttca atgtagggaa ttcgttcttg atggtttcct ccacagtttt tctccataat
     4201 cttgaagagg ccaaaacatt agctttatcc aaggtccaaa taggcaatgg tggctcatgt
     4261 tgtagggcca tgaaagcggc cattcttgtg attctttgca cttctggaac ggtgtattgt
     4321 tcactatccc aagcgacacc atcaccatcg tcttcctttc tcttaccaaa gtaaatacct
     4381 cccactaatt ctctgacaac aacgaagtca gtacctttag caaattgtgg cttgattgga
     4441 gataagtcta aaagagagtc ggatgcaaag ttacatggtc ttaagttggc gtacaattga
     4501 agttctttac ggatttttag taaaccttgt tcaggtctaa cactaccggt accccatttt
     4561 ggaccaccca cagcacctaa caaaacggca tcaaccttct tggaggcttc cagtgcctca
     4621 tctggaagtg ggacacctgt agcatcgata gcagcaccac caattaaatg attttcgaaa
     4681 tcgaacttga cattggaacg aacatcagaa atagctttaa gaaccttaat ggcttcggct
     4741 gtgatttctt gaccaacgtg gtcacctggc aaaacgacga tcttcttagg aacaaacatt
     4801 agaatggtat atccttgaaa tatatatata tattgctgaa atgtaaaagg taagaaaagt
     4861 tagaaagtaa gacgattgct aaccacctat tggaaaaaac aataggtcct taaataatat
     4921 tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct attctatatg
     4981 aaaagccggt tccggcctct cacctttcct ttttctccca atttttcagt tgaaaaaggt
     5041 atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg
     5101 tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag
     5161 attcgaactc ttgcatctta cgatacctga gtattcccac agttaacgaa gcatctgtgc
     5221 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     5281 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     5341 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     5401 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     5461 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     5521 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     5581 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     5641 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     5701 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     5761 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     5821 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     5881 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     5941 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     6001 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     6061 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     6121 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     6181 tttctagtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     6241 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6301 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6361 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6421 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6481 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6541 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6601 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6661 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6721 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6781 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6841 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6901 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6961 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7021 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7081 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7141 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7201 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7261 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7321 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7381 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7441 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7501 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7561 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7621 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7681 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7741 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7801 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7861 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7921 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7981 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8041 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8101 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8161 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8221 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8281 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8341 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     8401 cccgcgcgtt ggccgattca ttcccg
//
