LOCUS       40924_49722        5973 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector yGALset985, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  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 5973)
  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 5973)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5973)
  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..5973
                     /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    926..981
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     terminator      1036..1083
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1475..2821)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2935..3735)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3736..3951)
                     /label=URA3 promoter
     promoter        4067..4171
                     /label=AmpR promoter
     CDS             4172..5029
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5203..5791
                     /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 cgagctcgag atctgcagct ggtaccatgg
      961 aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1021 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1081 ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc
     1141 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     1201 ttacgcatct gtgcggtatt tcacaccgca tatatcggat cgtacttgtt acccatcatt
     1261 gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat ttgaacctgt
     1321 ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg ttcctggaga
     1381 aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc attttctgcg
     1441 tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc ttcattttgt
     1501 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     1561 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     1621 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     1681 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     1741 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     1801 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     1861 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     1921 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     1981 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     2041 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     2101 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     2161 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     2221 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     2281 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     2341 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     2401 gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     2461 gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa
     2521 atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt
     2581 gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta
     2641 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
     2701 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
     2761 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
     2821 cgatccgatg ataagctgtc aaacatgaga attgggtaat aactgatata attaaattga
     2881 agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg
     2941 ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac
     3001 cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt
     3061 agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa
     3121 acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct
     3181 ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt
     3241 agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc
     3301 tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc
     3361 caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga
     3421 gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa
     3481 attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt
     3541 caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc
     3601 cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg
     3661 catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata
     3721 tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt
     3781 taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct
     3841 aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt
     3901 tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga attgaaaagc
     3961 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4021 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4081 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4141 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5941 gcgcagcgag tcagtgagcg aggaagcgga aga
//
