LOCUS       40924_49702        6345 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset352, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  AUTHORS   Enomoto S, Chen G, Berman J.
  TITLE     Vectors for expressing T7 epitope- and His6 affinity-tagged fusion 
            proteins in S. cerevisiae
  JOURNAL   BioTechniques 24 (5), 782-786 (1998)
  PUBMED    9591127
REFERENCE   2  (bases 1 to 6345)
  AUTHORS   Enomoto S, Berman J.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-1998) Plant Biology, University of Minnesota, 220 
            Biological Sciences Center, St. Paul, MN 55108, USA
REFERENCE   3  (bases 1 to 6345)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6345)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "24"; issue: "5"; pages: 
            "782-786"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-JAN-1998) Plant Biology, University of Minnesota, 220 Biological
            Sciences Center, St. Paul, MN 55108, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..88
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(121..785)
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     RBS             804..826
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             846..863
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             867..899
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             903..926
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     misc_feature    936..982
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     terminator      1046..1093
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    complement(1807..2969)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     CDS             complement(3307..4107)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4108..4323)
                     /label=URA3 promoter
     promoter        4439..4543
                     /label=AmpR promoter
     CDS             4544..5401
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gatctcgatc
       61 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct aggatccggg
      121 gttttttctc cttgacgtta aagtatagag gtatattaac aattttttgt tgatactttt
      181 attacatttg aataagaagt aatacaaacc gaaaatgttg aaagtattag ttaaagtggt
      241 tatgcagttt ttgcatttat atatctgtta atagatcaaa aatcatcgct tcgctgatta
      301 attaccccag aaataaggct aaaaaactaa tcgcattatc atcctatggt tgttaatttg
      361 attcgttcat ttgaaggttt gtggggccag gttactgcca atttttcctc ttcataacca
      421 taaaagctag tattgtagaa tctttattgt tcggagcagt gcggcgcgag gcacatctgc
      481 gtttcaggaa cgcgaccggt gaagacgagg acgcacggag gagagtcttc cttcggaggg
      541 ctgtcacccg ctcggcggct tctaatccgt acttcaatat agcaatgagc agttaagcgt
      601 attactgaaa gttccaaaga gaaggttttt ttaggctaag ataatggggc tctttacatt
      661 tccacaacat ataagtaaga ttagatatgg atatgtatat ggatatgtat atggtggtaa
      721 tgccatgtaa tatgattatt aaacttcttt gcgtccatcc aaaaaaaaag taagaatttt
      781 tgaaaattcg aattctagat aattttgttt aactttaaga aggagatata catatgcggg
      841 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
      901 gggatctgta cgacgatgac gataaggatc gatggggatc cgagctcgag atctgcagct
      961 ggtaccatgg aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1021 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1081 gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc
     1141 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     1201 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatcgctg ggccattctc
     1261 atgaagaata tcttgaattt attgtcatat tactagttgg tgtggaagtc catatatcgg
     1321 tgatcaatat agtggttgac atgctggcta gtcaacattg agccttttga tcatgcaaat
     1381 atattacggt attttacaat caaatatcaa acttaactat tgactttata acttatttag
     1441 gtggtaacat tcttataaaa aagaaaaaaa ttactgcaaa acagtactag cttttaactt
     1501 gtatcctagg ttatctatgc tgtctcacca tagagaatat tacctatttc agaatgtatg
     1561 tccatgattc gccgggtaaa tacatataat acacaaatct ggcttaataa agtctataat
     1621 atatctcata aagaagtgct aaattggcta gtgctatata tttttaagaa aatttctttt
     1681 gactaagtcc atatcgactt tgtaaaagtt cactttagca tacatatatt acacgagcca
     1741 gaaattgtaa cttttgccta aaatcacaaa ttgcaaaatt taattgcttg caaaaggtca
     1801 catgcttata atcaactttt ttaaaaattt aaaatacttt tttatttttt atttttaaac
     1861 ataaatgaaa taatttattt attgtttatg attaccgaaa cataaaacct gctcaagaaa
     1921 aagaaactgt tttgtccttg gaaaaaaagc actacctagg agcggccaaa atgccgaggc
     1981 tttcatagct taaactcttt acagaaaata ggcattatag atcagttcga gttttcttat
     2041 tcttccttcc ggttttatcg tcacagtttt acagtaaata agtatcacct cttagagttc
     2101 gatgataagc tgtcaaacat gagaattaat tccacatgtt aaaatagtga aggagcatgt
     2161 tcggcacaca gtggaccgaa cgtggggtaa gtgcactagg gtccggttaa acggatctcg
     2221 cattgatgag gcaacgctaa ttatcaacat atagattgtt atctatctgc atgaacacga
     2281 aatctttact tgacgacttg aggctgatgg tgtttatgca aagaaaccac tgtgtttaat
     2341 atgtgtcact gtttgatatt actgtcagcg tagaagataa tagtaaaagc ggttaataag
     2401 tgtatttgag ataagtgtga taaagttttt acagcgaaaa gacgataaat acaagaaaat
     2461 gattacgagg atacggagag aggtatgtac atgtgtattt atatactaag ctgccggcgg
     2521 ttgtttgcaa gaccgagaaa aggctagcaa gaatcgggtc attgtagcgt atgcgcctgt
     2581 gaacattctc ttcaacaagt ttgattccat tgcggtgaaa tggtaaaagt caaccccctg
     2641 cgatgtatat tttcctgtac aatcaatcaa aaagccaaat gatttagcat tatctttaca
     2701 tcttgttatt ttacagattt tatgtttaga tcttttatgc ttgcttttca aaaggcttgc
     2761 aggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta tttcttagca
     2821 tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc cacacctccg
     2881 cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt ttctggcgtc
     2941 agtccaccag ctaacataaa atgtaagctc tcggggctct cttgccttcc aacccagtca
     3001 gaaatcgagt tccaatccaa aagttcacct gtcccacctg cttctgaatc aaacaaggga
     3061 ataaacgaat gaggtttctg tgaagctgca ctgagtagta tgttgcagtc ttttggaaat
     3121 acgagtcttt taataactgg caaaccgagg aactcttggt attcttgcca cgactcatct
     3181 ccatgcagtt ggacgatcga tgataagctg tcaaacatga gaattgggta ataactgata
     3241 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     3301 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     3361 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     3421 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     3481 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     3541 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     3601 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     3661 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     3721 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     3781 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     3841 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     3901 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     3961 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     4021 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     4081 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcatg tttttgttct
     4141 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     4201 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     4261 atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     4321 gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6241 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6301 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaga
//
