LOCUS       40924_49697        6852 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset351, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  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 6852)
  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 6852)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6852)
  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..6852
                     /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    934..980
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     terminator      1044..1091
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    complement(1805..2967)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     CDS             complement(3332..4423)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4436..4840)
                     /label=LEU2 promoter
     promoter        4946..5050
                     /label=AmpR promoter
     CDS             5051..5908
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6082..6670
                     /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 gatggatccg acctcgagat ctgcagctgg
      961 taccatggaa ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1021 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1081 ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg
     1141 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     1201 ttttctcctt acgcatctgt gcggtatttc acaccgcata tatcgctggg ccattctcat
     1261 gaagaatatc ttgaatttat tgtcatatta ctagttggtg tggaagtcca tatatcggtg
     1321 atcaatatag tggttgacat gctggctagt caacattgag ccttttgatc atgcaaatat
     1381 attacggtat tttacaatca aatatcaaac ttaactattg actttataac ttatttaggt
     1441 ggtaacattc ttataaaaaa gaaaaaaatt actgcaaaac agtactagct tttaacttgt
     1501 atcctaggtt atctatgctg tctcaccata gagaatatta cctatttcag aatgtatgtc
     1561 catgattcgc cgggtaaata catataatac acaaatctgg cttaataaag tctataatat
     1621 atctcataaa gaagtgctaa attggctagt gctatatatt tttaagaaaa tttcttttga
     1681 ctaagtccat atcgactttg taaaagttca ctttagcata catatattac acgagccaga
     1741 aattgtaact tttgcctaaa atcacaaatt gcaaaattta attgcttgca aaaggtcaca
     1801 tgcttataat caactttttt aaaaatttaa aatacttttt tattttttat ttttaaacat
     1861 aaatgaaata atttatttat tgtttatgat taccgaaaca taaaacctgc tcaagaaaaa
     1921 gaaactgttt tgtccttgga aaaaaagcac tacctaggag cggccaaaat gccgaggctt
     1981 tcatagctta aactctttac agaaaatagg cattatagat cagttcgagt tttcttattc
     2041 ttccttccgg ttttatcgtc acagttttac agtaaataag tatcacctct tagagttcga
     2101 tgataagctg tcaaacatga gaattaattc cacatgttaa aatagtgaag gagcatgttc
     2161 ggcacacagt ggaccgaacg tggggtaagt gcactagggt ccggttaaac ggatctcgca
     2221 ttgatgaggc aacgctaatt atcaacatat agattgttat ctatctgcat gaacacgaaa
     2281 tctttacttg acgacttgag gctgatggtg tttatgcaaa gaaaccactg tgtttaatat
     2341 gtgtcactgt ttgatattac tgtcagcgta gaagataata gtaaaagcgg ttaataagtg
     2401 tatttgagat aagtgtgata aagtttttac agcgaaaaga cgataaatac aagaaaatga
     2461 ttacgaggat acggagagag gtatgtacat gtgtatttat atactaagct gccggcggtt
     2521 gtttgcaaga ccgagaaaag gctagcaaga atcgggtcat tgtagcgtat gcgcctgtga
     2581 acattctctt caacaagttt gattccattg cggtgaaatg gtaaaagtca accccctgcg
     2641 atgtatattt tcctgtacaa tcaatcaaaa agccaaatga tttagcatta tctttacatc
     2701 ttgttatttt acagatttta tgtttagatc ttttatgctt gcttttcaaa aggcttgcag
     2761 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     2821 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     2881 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     2941 tccaccagct aacataaaat gtaagctctc ggggctctct tgccttccaa cccagtcaga
     3001 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     3061 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     3121 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     3181 atgcagttgg acgatcgatg ataagctgtc aaacatgaga attaattcta ccctatgaac
     3241 atattccatt ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt
     3301 acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga
     3361 cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg
     3421 catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg
     3481 acaatttcaa catcattgca gcagacaaga tagtggcgat agggtcaacc ttattctttg
     3541 gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg
     3601 gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt
     3661 catcggagat gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg
     3721 ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag
     3781 gaaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa
     3841 cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag
     3901 cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga
     3961 caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga
     4021 caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag
     4081 agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt
     4141 ttagtaaacc ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac
     4201 ctaacaaaac ggcatcaacc ttcttggagg cttccagcgc ctcatctgga agtgggacac
     4261 ctgtagcatc gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg
     4321 aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa
     4381 cgtggtcacc tggcaaaacg acgatcttct taggggcaga cataggggca gacattagaa
     4441 tggtatatcc ttgaaatata tatatatatt gctgaaatgt aaaaggtaag aaaagttaga
     4501 aagtaagacg attgctaacc acctattgga aaaaacaata ggtccttaaa taatattgtc
     4561 aacttcaagt attgtgatgc aagcatttag tcatgaacgc ttctctattc tatatgaaaa
     4621 gccggttccg gcctctcacc tttccttttt ctcccaattt ttcagttgaa aaaggtatat
     4681 gcgtcaggcg acctctgaaa ttaacaaaaa atttccagtc atcgaatttg attctgtgcg
     4741 atagcgcccc tgtgtgttct cgttatgttg aggaaaaaaa taatggttgc taagagattc
     4801 gaactcttgc atcttacgat acctgagtat tcccacagtt aattcttgaa gacgaaaggg
     4861 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4921 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4981 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5041 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5101 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5161 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5221 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5281 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5341 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5401 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5461 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5521 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5581 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5641 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5701 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5761 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5821 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5881 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5941 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6001 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6841 ggaagcggaa ga
//
