LOCUS       40924_35364        9690 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pPP1MFmds1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9690)
  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  JOURNAL   Unpublished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 9690)
  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 9690)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9690)
  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            "Unpublished"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..9690
                     /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             1228..2727
                     /codon_start=1
                     /note="unnamed protein product; mds1 mature sequence"
                     /protein_id="SJL87755.1"
                     /translation="ATKRGSPNPTRAAAVKAAFQTSWNAYHHFAFPHDDLHPVSNSFDD
                     ERNGWGSSAIDGLDTAILMGDADIVNTILQYVPQINFTTTAVANQGISVFETNIRYLGG
                     LLSAYDLLRGPFSSLATNQTLVNSLLRQAQTLANGLKVAFTTPSGVPDPTVFFNPTVRR
                     SGASSNNVAEIGSLVLEWTRLSDLTGNPQYAQLAQKGESYLLNPKGSPEAWPGLIGTFV
                     STSNGTFQDSSGSWSGLMDSFYEYLIKMYLYDPVAFAHYKDRWVLGADSTIGHLGSHPS
                     TRKDLTFLSSYNGQSTSPNSGHLASFGGGNFILGGILLNEQKYIDFGIKLASSYFGTYT
                     QTASGIGPEGFAWVDSVTGAGGSPPSSQSGFYSSAGFWVTAPYYILRPETLESLYYAYR
                     VTGDSKWQDLAWEALSAIEDACRAGSAYSSINDVTQANGGGASDDMESFWFAEALKYAY
                     LIFAEESDVQVQATGGNKFVFNTEAHPFSIRSSSRRGGHLA"
     terminator      2988..3234
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(3647..6181)
                     /codon_start=1
                     /note="unnamed protein product; HIS4"
                     /protein_id="SJL87756.1"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     misc_feature    6536..7292
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     misc_feature    7435..7575
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7761..8349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8523..9380)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9381..9485)
                     /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 aagcttacgt agaattcgcc acaaaacgtg gatctcccaa ccctacgagg
     1261 gcggcagcag tcaaggccgc attccagacg tcgtggaacg cttaccacca ttttgccttt
     1321 ccccatgacg acctccaccc ggtcagcaac agctttgatg atgagagaaa cggctggggc
     1381 tcgtcggcaa tcgatggctt ggacacggct atcctcatgg gggatgccga cattgtgaac
     1441 acgatccttc agtatgtacc gcagatcaac ttcaccacga ctgcggttgc caaccaaggc
     1501 atctccgtgt tcgagaccaa cattcggtac ctcggtggcc tgctttctgc ctatgacctg
     1561 ttgcgaggtc ctttcagctc cttggcgaca aaccagaccc tggtaaacag ccttctgagg
     1621 caggctcaaa cactggccaa cggcctcaag gttgcgttca ccactcccag cggtgtcccg
     1681 gaccctaccg tcttcttcaa ccctactgtc cggagaagtg gtgcatctag caacaacgtc
     1741 gctgaaattg gaagcctggt gctcgagtgg acacggttga gcgacctgac gggaaacccg
     1801 cagtatgccc agcttgcgca gaagggcgag tcgtatctcc tgaatccaaa gggaagcccg
     1861 gaggcatggc ctggcctgat tggaacgttt gtcagcacga gcaacggtac ctttcaggat
     1921 agcagcggca gctggtccgg cctcatggac agcttctacg agtacctgat caagatgtac
     1981 ctgtacgacc cggttgcgtt tgcacactac aaggatcgct gggtccttgg tgccgactcg
     2041 accattgggc atctcggctc tcacccgtcg acgcgcaagg acttgacctt tttgtcttcg
     2101 tacaacggac agtctacgtc gccaaactca ggacatttgg ccagttttgg cggtggcaac
     2161 ttcatcttgg gaggcattct cctgaacgag caaaagtaca ttgactttgg aatcaagctt
     2221 gccagctcgt actttggcac gtacacccag acggcttctg gaatcggccc cgaaggcttc
     2281 gcgtgggtgg acagcgtgac gggcgccggc ggctcgccgc cctcgtccca gtccgggttc
     2341 tactcgtcgg caggattctg ggtgacggca ccgtattaca tcctgcggcc ggagacgctg
     2401 gagagcttgt actacgcata ccgcgtcacg ggcgactcca agtggcagga cctggcgtgg
     2461 gaagcgttga gtgccattga ggacgcatgc cgcgccggca gcgcgtactc gtccatcaac
     2521 gacgtgacgc aggccaacgg cgggggtgcc tctgacgata tggagagctt ctggtttgcc
     2581 gaggcgctca agtatgcgta cctgatcttt gcggaggagt cggatgtgca ggtgcaggcc
     2641 accggcggga acaaatttgt ctttaacacg gaggcgcacc cctttagcat ccgttcatca
     2701 tcacgacggg gcggccacct tgcttaaagg cgctgagagt tctctcgcct gtttttttgc
     2761 tcgtattatt cattttctgc tgctcgtccc gaatcgctcc gtagatattt tgacacatgc
     2821 gtctggatag agcggctggg accaccgaat atggtatgtc attcgagata tgaagcgcaa
     2881 gagatccggg ccctctagat gcggccgcga attaattcgc cttagacatg actgttcctc
     2941 agttcaagtt gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc
     3001 agaatgccat ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct
     3061 atatagtata ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca
     3121 gcctatctcg cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg
     3181 atgtttttct tggtatttcc cactcctctt cagagtacag aagattaagt gagaagttcg
     3241 tttgtgcaag cttatcgata agctttaatg cggtagttta tcacagttaa attgctaacg
     3301 cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac
     3361 cctggatgct gtaggcatag gcttggttat gccggtactg ccgggcctct tgcgggatat
     3421 cgtccattcc gacagcatcg ccagtcacta tggcgtgctg ctagcgctat atgcgttgat
     3481 gcaatttcta tgcgcacccg ttctcggagc actgtccgac cgctttggcc gccgcccagt
     3541 cctgctcgct tcgctacttg gagccactat cgactacgcg atcatggcga ccacacccgt
     3601 cctgtggatc tatcgaatct aaatgtaagt taaaatctct aaataattaa ataagtccca
     3661 gtttctccat acgaacctta acagcattgc ggtgagcatc tagaccttca acagcagcca
     3721 gatccatcac tgcttggcca atatgtttca gtccctcagg agttacgtct tgtgaagtga
     3781 tgaacttctg gaaggttgca gtgttaactc cgctgtattg acgggcatat ccgtacgttg
     3841 gcaaagtgtg gttggtaccg gaggagtaat ctccacaact ctctggagag taggcaccaa
     3901 caaacacaga tccagcgtgt tgtacttgat caacataaga agaagcattc tcgatttgca
     3961 ggatcaagtg ttcaggagcg tactgattgg acatttccaa agcctgctcg taggttgcaa
     4021 ccgatagggt tgtagagtgt gcaatacact tgcgtacaat ttcaaccctt ggcaactgca
     4081 cagcttggtt gtgaacagca tcttcaattc tggcaagctc cttgtctgtc atatcgacag
     4141 ccaacagaat cacctgggaa tcaataccat gttcagcttg agacagaagg tctgaggcaa
     4201 cgaaatctgg atcagcgtat ttatcagcaa taactagaac ttcagaaggc ccagcaggca
     4261 tgtcaatact acacagggct gatgtgtcat tttgaaccat catcttggca gcagtaacga
     4321 actggtttcc tggaccaaat attttgtcac acttaggaac agtttctgtt ccgtaagcca
     4381 tagcagctac tgcctgggcg cctcctgcta gcacgataca cttagcacca accttgtggg
     4441 caacgtagat gacttctggg gtaagggtac catccttctt aggtggagat gcaaaaacaa
     4501 tttctttgca accagcaact ttggcaggaa cacccagcat cagggaagtg gaaggcagaa
     4561 ttgcggttcc accaggaata tagaggccaa ctttctcaat aggtcttgca aaacgagagc
     4621 agactacacc agggcaagtc tcaacttgca acgtctccgt tagttgagct tcatggaatt
     4681 tcctgacgtt atctatagag agatcaatgg ctctcttaac gttatctggc aattgcataa
     4741 gttcctctgg gaaaggagct tctaacacag gtgtcttcaa agcgactcca tcaaacttgg
     4801 cagttagttc taaaagggct ttgtcaccat tttgacgaac attgtcgaca attggtttga
     4861 ctaattccat aatctgttcc gttttctgga taggacgacg aagggcatct tcaatttctt
     4921 gtgaggaggc cttagaaacg tcaattttgc acaattcaat acgaccttca gaagggactt
     4981 ctttaggttt ggattcttct ttaggttgtt ccttggtgta tcctggcttg gcatctcctt
     5041 tccttctagt gacctttagg gacttcatat ccaggtttct ctccacctcg tccaacgtca
     5101 caccgtactt ggcacatcta actaatgcaa aataaaataa gtcagcacat tcccaggcta
     5161 tatcttcctt ggatttagct tctgcaagtt catcagcttc ctccctaatt ttagcgttca
     5221 acaaaacttc gtcgtcaaat aaccgtttgg tataagaacc ttctggagca ttgctcttac
     5281 gatcccacaa ggtggcttcc atggctctaa gaccctttga ttggccaaaa caggaagtgc
     5341 gttccaagtg acagaaacca acacctgttt gttcaaccac aaatttcaag cagtctccat
     5401 cacaatccaa ttcgataccc agcaactttt gagttgctcc agatgtagca cctttatacc
     5461 acaaaccgtg acgacgagat tggtagactc cagtttgtgt ccttatagcc tccggaatag
     5521 actttttgga cgagtacacc aggcccaacg agtaattaga agagtcagcc accaaagtag
     5581 tgaatagacc atcggggcgg tcagtagtca aagacgccaa caaaatttca ctgacaggga
     5641 actttttgac atcttcagaa agttcgtatt cagtagtcaa ttgccgagca tcaataatgg
     5701 ggattatacc agaagcaaca gtggaagtca catctaccaa ctttgcggtc tcagaaaaag
     5761 cataaacagt tctactaccg ccattagtga aacttttcaa atcgcccagt ggagaagaaa
     5821 aaggcacagc gatactagca ttagcgggca aggatgcaac tttatcaacc agggtcctat
     5881 agataaccct agcgcctggg atcatccttt ggacaactct ttctgccaaa tctaggtcca
     5941 aaatcacttc attgatacca ttattgtaca acttgagcaa gttgtcgatc agctcctcaa
     6001 attggtcctc tgtaacggat gactcaactt gcacattaac ttgaagctca gtcgattgag
     6061 tgaacttgat caggttgtgc agctggtcag cagcataggg aaacacggct tttcctacca
     6121 aactcaagga attatcaaac tctgcaacac ttgcgtatgc aggtagcaag ggaaatgtca
     6181 tacttgaagt cggacagtga gtgtagtctt gagaaattct gaagccgtat ttttattatc
     6241 agtgagtcag tcatcaggag atcctctacg ccggacgcat cgtggccggc atcaccggcg
     6301 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     6361 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     6421 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     6481 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
     6541 tatctatgat tggaagtatg ggaatggtga tacccgcatt cttcagtgtc ttgaggtctc
     6601 ctatcagatt atgcccaact aaagcaaccg gaggaggaga tttcatggta aatttctctg
     6661 acttttggtc atcagtagac tcgaactgtg agactatctc ggttatgaca gcagaaatgt
     6721 ccttcttgga gacagtaaat gaagtcccac caataaagaa atccttgtta tcaggaacaa
     6781 acttcttgtt tcgaactttt tcggtgcctt gaactataaa atgtagagtg gatatgtcgg
     6841 gtaggaatgg agcgggcaaa tgcttacctt ctggaccttc aagaggtatg tagggtttgt
     6901 agatactgat gccaacttca gtgacaacgt tgctatttcg ttcaaaccat tccgaatcca
     6961 gagaaatcaa agttgtttgt ctactattga tccaagccag tgcggtcttg aaactgacaa
     7021 tagtgtgctc gtgttttgag gtcatctttg tatgaataaa tctagtcttt gatctaaata
     7081 atcttgacga gccaaggcga taaataccca aatctaaaac tcttttaaaa cgttaaaagg
     7141 acaagtatgt ctgcctgtat taaaccccaa atcagctcgt agtctgatcc tcatcaactt
     7201 gaggggcact atcttgtttt agagaaattt gcggagatgc gatatcgaga aaaaggtacg
     7261 ctgattttaa acgtgaaatt tatctcaaga tctctgcctc gcgcgtttcg gtgatgacgg
     7321 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7381 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     7441 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     7501 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7561 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8161 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8821 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9121 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9481 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     9541 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt aattctcatg
     9601 tttgacagct tatcatcgat aagctgactc atgttggtat tgtgaaatag acgcagatcg
     9661 ggaacactga aaaataacag ttattattcg
//
