LOCUS       .                       6038 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             733..750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             522..539
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     promoter        475..493
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     CDS             579..602
                     /codon_start=1
                     /product="Xpress™ epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /transl_table=1
                     /translation="DLY,DDDDK"
                     /label="Xpress™ tag"
                     /note="color: #cc99b2"
     CDS             543..575
                     /codon_start=1
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /transl_table=1
                     /translation="MASMTGGQQMG"
                     /label="T7 tag (gene 10 leader)"
     CDS             682..723
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
                     /note="color: #cc99b2"
     protein_bind    2..119
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
                     /note="color: #31849b"
                     /label="UAS"
     promoter        complement(3798..4013)
                     /gene="S. cerevisiae URA3"
                     /label="URA3 promoter"
                     /note="color: #ffffff"
     terminator      784..1031
                     /gene="S. cerevisiae CYC1"
                     /note="transcription terminator for CYC1"
                     /note="color: #ffffff"
                     /label="CYC1 terminator"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /note="inducible promoter, regulated by Gal4"
                     /note="color: #ffffff"
                     /label="GAL1 promoter"
     rep_origin      complement(5566..6021)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     rep_origin      complement(1279..1867)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     CDS             complement(2994..3797)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /note="yeast auxotrophic marker, counterselectable with
                     5-fluoroorotic acid (5-FOA)"
                     /note="color: #ff7f50"
                     /product="orotidine-5'-phosphate decarboxylase, required
                     for uracil biosynthesis"
                     /transl_table=1
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN*"
                     /label="URA3"
     CDS             complement(2038..2898)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance fo ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      4617..5497
                     /note="yeast 2μ plasmid origin of replication"
                     /note="color: #ffff00"
                     /label="2μ ori"
     CDS             510..512
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     misc_feature    602..669
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagcttacca tggggggttc tcatcatcat catcatcatg
      541 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      601 aggtacctag gatccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgctc
      661 gagtctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
      721 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc ctagagggcc
      781 gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
      841 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
      901 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
      961 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     1021 ctttaatttg caagctgcgg ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1081 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1141 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1201 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aagcccagga accgtaaaaa
     1261 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1321 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1381 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1441 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1501 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1561 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1621 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1681 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1741 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1801 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1861 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1921 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1981 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2041 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2101 tgcctgactc cccgtcgtgt agataactac gatacgggag cgcttaccat ctggccccag
     2161 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2221 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2281 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2341 tgttggcatt gctacaggca tcgtggtgtc actctcgtcg tttggtatgg cttcattcag
     2401 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2461 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2521 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2581 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2641 ttgcccggcg tcaatacggg ataatagtgt atcacatagc agaactttaa aagtgctcat
     2701 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2761 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2821 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2881 gaaatgttga atactcatac tcttcctttt tcaatgggta ataactgata taattaaatt
     2941 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     3001 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     3061 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     3121 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     3181 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     3241 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     3301 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     3361 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     3421 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     3481 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     3541 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     3601 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     3661 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     3721 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     3781 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
     3841 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     3901 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     3961 tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa
     4021 gctagcttat cgatgataag ctgtcaaaga tgagaattaa ttccacggac tatagactat
     4081 actagatact ccgtctactg tacgatacac ttccgctcag gtccttgtcc tttaacgagg
     4141 ccttaccact cttttgttac tctattgatc cagctcagca aaggcagtgt gatctaagat
     4201 tctatcttcg cgatgtagta aaactagcta gaccgagaaa gagactagaa atgcaaaagg
     4261 cacttctaca atggctgcca tcattattat ccgatgtgac gctgcagctt ctcaatgata
     4321 ttcgaatacg ctttgaggag atacagccta atatccgaca aactgtttta cagatttacg
     4381 atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt ttccctgaaa
     4441 cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg gaagacaatg
     4501 tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc ttgcacgtcg
     4561 catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc tttgttaacg
     4621 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     4681 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     4741 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgacga gagcgctaat
     4801 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct
     4861 attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg
     4921 ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc
     4981 agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt
     5041 gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag
     5101 cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg
     5161 tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga
     5221 aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat
     5281 tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag
     5341 agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga
     5401 gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat
     5461 acttttgagc aatgtttgtg gaagcggtat tcgcaatggg aagctccacc ccggttgata
     5521 atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaaacgtta
     5581 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aacgaatagc
     5641 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5701 ttccagtttc caacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5761 aaagggtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5821 ggtcgaggtg ccgtaaagca gtaaatcgga agggtaaacg gatgccccca tttagagctt
     5881 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcggggg
     5941 ctagggcggt gggaagtgta ggggtcacgc tgggcgtaac caccacaccc gccgcgctta
     6001 atggggcgct acagggcgcg tggggatgat ccactagt
//
