LOCUS       40924_35354        9568 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pPP1IVHAsi, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9568)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9568)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 9568)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9568)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..937
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             949..1215
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
     CDS             1510..1536
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     terminator      2854..3100
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(3513..6047)
                     /codon_start=1
                     /note="unnamed protein product; HIS4"
                     /protein_id="SJL88005.1"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     misc_feature    6414..7170
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     misc_feature    7313..7453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7639..8227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8401..9258)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9259..9363)
                     /label=AmpR promoter
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcgcca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaagga tccaaacgat gagatttcct
      961 tcaattttta ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact
     1021 acaacagaag atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta
     1081 gaaggggatt tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg
     1141 tttataaata ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa
     1201 agagaggctg aagcccaaga ccttccagga aatgacaaca acagcacagc aacgctgtgc
     1261 ctgggacatc atgcggtgcc aaacggaacg ctagtgaaaa caatcacgaa tgatcagatt
     1321 gaagtgacta atgctactga gctggttcag agttcctcaa cgggtaaaat atgcaacaat
     1381 cctcatcgaa tccttgatgg aataaactgc acactgatag atgctctatt gggggaccct
     1441 cattgtgatg gatttcaaaa tgagaaatgg gaccttttcg ttgaacgcag caaagctttc
     1501 agcaactgtt acccttatga tgtgccagat tatgcctccc ttaggtcact agttgcctcg
     1561 tcaggcactc tggagtttat caatgaaggc ttcaattgga ctggggtcac tcagaatggg
     1621 ggaagcagtg cttgcaaaag aggacctgat agcggttttt tcagtagact gaactggttg
     1681 tacaaatcag gaagcacata tccagtgcaa aacgtgacca tgccaaacaa tgacaattct
     1741 gacaaactat acatttgggg ggttcaccac ccgagcacgg acaaagaaca aaccaaccta
     1801 tatgttcaag catcagggaa agtcacagtc tccaccaaga gaagccagca aactataatc
     1861 ccgaatgtcg ggtctagacc ctgggtaagg ggtctgtcta gtagaataag catctattgg
     1921 acaatagtta aaccgggaga catactggta attaatagta atgggaacct aattgctcct
     1981 cggggttact tcaaaatgcg cactgggaaa agctcaataa tgaggtcaga tgcacctatt
     2041 ggcacctgca gctctgaatg catcactcca aatggaagca ttcccaatga caagcccttt
     2101 caaaacgtaa acaagatcac atatggggca tgtcccaagt atgttaagca aaacactctg
     2161 aagttggcaa cagggatgcg gaatgtacca gagaaacaaa ctagaggcat attcggcgca
     2221 atagcaggtt tcatagaaaa tggttgggag ggaatgatag acggttggta cggtttcagg
     2281 catcaaaatt ccgagggcac aggacaagca gcagatctta aaagcactca agcagccatc
     2341 gaccaaatca atgggaaact gaatagggta atcgagaaga cgaacgagaa attccatcaa
     2401 atcgaaaagg aattctcaga agtagaaggg agaattcagg acctcgagaa atacgttgaa
     2461 gacactaaaa tagatctctg gtcttacaat gcggagcttc ttgtcgctct ggagaaccaa
     2521 catacaattg atctgactga ctcggaaatg aacaaactgt ttgaaaaaac aaggaggcaa
     2581 ctgagggaaa atgctgagga catgggcaat ggttgcttca aaatatacca caaatgtgac
     2641 aatgcttgca tagggtcaat cagaaatggg acttatgacc atgatgtata cagagacgaa
     2701 gcattaaaca accggtttca gatcaaaggt gttgaactga agtcaggata caaagactgg
     2761 atctaagcgg ccgcgaatta attcgcctta gacatgactg ttcctcagtt caagttgggc
     2821 acttacgaga agaccggtct tgctagattc taatcaagag gatgtcagaa tgccatttgc
     2881 ctgagagatg caggcttcat ttttgatact tttttatttg taacctatat agtataggat
     2941 tttttttgtc attttgtttc ttctcgtacg agcttgctcc tgatcagcct atctcgcagc
     3001 tgatgaatat cttgtggtag gggtttggga aaatcattcg agtttgatgt ttttcttggt
     3061 atttcccact cctcttcaga gtacagaaga ttaagtgaga agttcgtttg tgcaagctta
     3121 tcgataagct ttaatgcggt agtttatcac agttaaattg ctaacgcagt caggcaccgt
     3181 gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag
     3241 gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc cattccgaca
     3301 gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa tttctatgcg
     3361 cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc
     3421 tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg tggatctatc
     3481 gaatctaaat gtaagttaaa atctctaaat aattaaataa gtcccagttt ctccatacga
     3541 accttaacag cattgcggtg agcatctaga ccttcaacag cagccagatc catcactgct
     3601 tggccaatat gtttcagtcc ctcaggagtt acgtcttgtg aagtgatgaa cttctggaag
     3661 gttgcagtgt taactccgct gtattgacgg gcatatccgt acgttggcaa agtgtggttg
     3721 gtaccggagg agtaatctcc acaactctct ggagagtagg caccaacaaa cacagatcca
     3781 gcgtgttgta cttgatcaac ataagaagaa gcattctcga tttgcaggat caagtgttca
     3841 ggagcgtact gattggacat ttccaaagcc tgctcgtagg ttgcaaccga tagggttgta
     3901 gagtgtgcaa tacacttgcg tacaatttca acccttggca actgcacagc ttggttgtga
     3961 acagcatctt caattctggc aagctccttg tctgtcatat cgacagccaa cagaatcacc
     4021 tgggaatcaa taccatgttc agcttgagac agaaggtctg aggcaacgaa atctggatca
     4081 gcgtatttat cagcaataac tagaacttca gaaggcccag caggcatgtc aatactacac
     4141 agggctgatg tgtcattttg aaccatcatc ttggcagcag taacgaactg gtttcctgga
     4201 ccaaatattt tgtcacactt aggaacagtt tctgttccgt aagccatagc agctactgcc
     4261 tgggcgcctc ctgctagcac gatacactta gcaccaacct tgtgggcaac gtagatgact
     4321 tctggggtaa gggtaccatc cttcttaggt ggagatgcaa aaacaatttc tttgcaacca
     4381 gcaactttgg caggaacacc cagcatcagg gaagtggaag gcagaattgc ggttccacca
     4441 ggaatataga ggccaacttt ctcaataggt cttgcaaaac gagagcagac tacaccaggg
     4501 caagtctcaa cttgcaacgt ctccgttagt tgagcttcat ggaatttcct gacgttatct
     4561 atagagagat caatggctct cttaacgtta tctggcaatt gcataagttc ctctgggaaa
     4621 ggagcttcta acacaggtgt cttcaaagcg actccatcaa acttggcagt tagttctaaa
     4681 agggctttgt caccattttg acgaacattg tcgacaattg gtttgactaa ttccataatc
     4741 tgttccgttt tctggatagg acgacgaagg gcatcttcaa tttcttgtga ggaggcctta
     4801 gaaacgtcaa ttttgcacaa ttcaatacga ccttcagaag ggacttcttt aggtttggat
     4861 tcttctttag gttgttcctt ggtgtatcct ggcttggcat ctcctttcct tctagtgacc
     4921 tttagggact tcatatccag gtttctctcc acctcgtcca acgtcacacc gtacttggca
     4981 catctaacta atgcaaaata aaataagtca gcacattccc aggctatatc ttccttggat
     5041 ttagcttctg caagttcatc agcttcctcc ctaattttag cgttcaacaa aacttcgtcg
     5101 tcaaataacc gtttggtata agaaccttct ggagcattgc tcttacgatc ccacaaggtg
     5161 gcttccatgg ctctaagacc ctttgattgg ccaaaacagg aagtgcgttc caagtgacag
     5221 aaaccaacac ctgtttgttc aaccacaaat ttcaagcagt ctccatcaca atccaattcg
     5281 atacccagca acttttgagt tgctccagat gtagcacctt tataccacaa accgtgacga
     5341 cgagattggt agactccagt ttgtgtcctt atagcctccg gaatagactt tttggacgag
     5401 tacaccaggc ccaacgagta attagaagag tcagccacca aagtagtgaa tagaccatcg
     5461 gggcggtcag tagtcaaaga cgccaacaaa atttcactga cagggaactt tttgacatct
     5521 tcagaaagtt cgtattcagt agtcaattgc cgagcatcaa taatggggat tataccagaa
     5581 gcaacagtgg aagtcacatc taccaacttt gcggtctcag aaaaagcata aacagttcta
     5641 ctaccgccat tagtgaaact tttcaaatcg cccagtggag aagaaaaagg cacagcgata
     5701 ctagcattag cgggcaagga tgcaacttta tcaaccaggg tcctatagat aaccctagcg
     5761 cctgggatca tcctttggac aactctttct gccaaatcta ggtccaaaat cacttcattg
     5821 ataccattat tgtacaactt gagcaagttg tcgatcagct cctcaaattg gtcctctgta
     5881 acggatgact caacttgcac attaacttga agctcagtcg attgagtgaa cttgatcagg
     5941 ttgtgcagct ggtcagcagc atagggaaac acggcttttc ctaccaaact caaggaatta
     6001 tcaaactctg caacacttgc gtatgcaggt agcaagggaa atgtcatact tgaagtcgga
     6061 cagtgagtgt agtcttgaga aattctgaag ccgtattttt attatcagtg agtcagtcat
     6121 caggagatcc tctacgccgg acgcatcgtg gccgacctgc aggtcggcat caccggcgcc
     6181 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     6241 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     6301 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     6361 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagta
     6421 tctatgattg gaagtatggg aatggtgata cccgcattct tcagtgtctt gaggtctcct
     6481 atcagattat gcccaactaa agcaaccgga ggaggagatt tcatggtaaa tttctctgac
     6541 ttttggtcat cagtagactc gaactgtgag actatctcgg ttatgacagc agaaatgtcc
     6601 ttcttggaga cagtaaatga agtcccacca ataaagaaat ccttgttatc aggaacaaac
     6661 ttcttgtttc gaactttttc ggtgccttga actataaaat gtagagtgga tatgtcgggt
     6721 aggaatggag cgggcaaatg cttaccttct ggaccttcaa gaggtatgta gggtttgtag
     6781 atactgatgc caacttcagt gacaacgttg ctatttcgtt caaaccattc cgaatccaga
     6841 gaaatcaaag ttgtttgtct actattgatc caagccagtg cggtcttgaa actgacaata
     6901 gtgtgctcgt gttttgaggt catctttgta tgaataaatc tagtctttga tctaaataat
     6961 cttgacgagc caaggcgata aatacccaaa tctaaaactc ttttaaaacg ttaaaaggac
     7021 aagtatgtct gcctgtatta aaccccaaat cagctcgtag tctgatcctc atcaacttga
     7081 ggggcactat cttgttttag agaaatttgc ggagatgcga tatcgagaaa aaggtacgct
     7141 gattttaaac gtgaaattta tctcaagatc tctgcctcgc gcgtttcggt gatgacggtg
     7201 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     7261 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     7321 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     7381 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7441 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7501 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7801 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8041 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8221 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8341 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8521 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8701 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8821 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9001 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9361 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     9421 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattaa ttctcatgtt
     9481 tgacagctta tcatcgataa gctgactcat gttggtattg tgaaatagac gcagatcggg
     9541 aacactgaaa aataacagtt attattcg
//
