LOCUS       40924_49652        4989 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Yeast episomal plasmid with a TRP1 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Gietz RD, Sugino A.
  TITLE     New yeast-Escherichia coli shuttle vectors constructed with in vitro
            mutagenized yeast genes lacking six-base pair restriction sites.
  JOURNAL   Gene 1988;74:527-34.
  PUBMED    3073106
REFERENCE   2  (bases 1 to 4989)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4989)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1988"; volume: "74"; pages: "527-34"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     The sequence of this plasmid was reconstructed using the description
            from Gietz and Sugino.
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(233..289)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(743..2089)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2204..2875)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2876..2977)
                     /label=TRP1 promoter
     promoter        3083..3187
                     /label=AmpR promoter
     CDS             3188..4045
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4219..4807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
      481 gcggtatttc acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca
      541 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat
      601 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc
      661 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc
      721 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg
      781 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
      841 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
      901 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
      961 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     1021 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     1081 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     1141 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     1201 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     1261 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     1321 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     1381 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     1441 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     1501 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     1561 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     1621 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag
     1681 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     1741 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     1801 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     1861 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     1921 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     1981 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     2041 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat
     2101 aagctgtcaa acatgagaat tgatctttta tgcttgcttt tcaaaaggct tgcaggcaag
     2161 tgcacaaaca atacttaaat aaatactact cagtaataac ctatttctta gcatttttga
     2221 cgaaatttgc tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat
     2281 caacaccaat aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac
     2341 cagctaacat aaaatgtaag ctctcggggc tctcttgcct tccaacccag tcagaaatcg
     2401 agttccaatc caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg
     2461 aatgaggttt ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc
     2521 ttttaataac tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca
     2581 gttggacgat atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat
     2641 tacgaaacac gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa
     2701 gacttgaaat tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata
     2761 taatacccag caagtcagca tcggaatcta gtgcacattc tgcggcctct gtgctctgca
     2821 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     2881 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     2941 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcttgaagac gaaagggcct
     3001 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3181 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3301 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3361 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3421 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3481 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3601 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3721 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3781 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4021 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4081 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4141 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4321 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4921 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4981 agcggaaga
//
