LOCUS       40924_49712        6483 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset983, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6483)
  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 6483)
  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 6483)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6483)
  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..6483
                     /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"
     rep_origin      complement(1485..2831)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2972..4012)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="KITAEAIKVLKAISDVRSNVKFDFENHLIGGAAIDATGVPLPDEA
                     LEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYANLRPCNFASDSLLDLSP
                     IKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYTVPEVQRITRMAAFMALQ
                     HEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQLIDSAAMILVKNPTHLN
                     GIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAFGLYEPCHGSAPDLPKNK
                     VDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTGDLGGSNSTTEVGDAVAE
                     EVKKILA"
     promoter        complement(4074..4471)
                     /label=LEU2 promoter
     promoter        4577..4681
                     /label=AmpR promoter
     CDS             4682..5539
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5713..6301
                     /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 tatatcggat cgtacttgtt
     1261 acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat
     1321 ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg
     1381 ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc
     1441 attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc
     1501 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     1561 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     1621 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     1681 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag
     1741 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa
     1801 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact
     1861 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc
     1921 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc
     1981 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact
     2041 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt
     2101 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc
     2161 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat
     2221 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg
     2281 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt
     2341 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt
     2401 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac
     2461 tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     2521 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     2581 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     2641 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     2701 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     2761 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     2821 gatattggat cgatccgatg ataagctgtc aaacatgaga attaattcta ccctatgaac
     2881 atattccatt ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt
     2941 acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga
     3001 cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg
     3061 catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg
     3121 acaatttcaa catcattgca gcagacaaga tagtggcgat agggtcaacc ttattctttg
     3181 gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg
     3241 gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt
     3301 catcggagat gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg
     3361 ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag
     3421 gaaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa
     3481 cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag
     3541 cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga
     3601 caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga
     3661 caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag
     3721 agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt
     3781 ttagtaaacc ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac
     3841 ctaacaaaac ggcatcaacc ttcttggagg cttccagcgc ctcatctgga agtgggacac
     3901 ctgtagcatc gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg
     3961 aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatc ttgaccaacg
     4021 tggtcacctg gcaaaacgac gatcttctta ggggcagaca taggggcaga cattagaatg
     4081 tatatccttg aaatatatat atatattgct gaaatgtaaa aggtaagaaa agttagaata
     4141 agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt
     4201 caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg
     4261 ttccggcctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgcg
     4321 gcgacctctg aaattaacaa aaaatttcca gtcatcgaat ttgtctgtgc gatagcgccc
     4381 ctgtgtgttc tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt cgaactcttg
     4441 catcttacga tacctgagta ttcccacagt taattcttga agacgaaagg gcctcgtgat
     4501 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4561 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4621 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4681 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4801 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4861 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4921 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4981 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5101 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5221 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5281 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5521 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5821 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6481 aga
//
