LOCUS       sequence-251549-        8344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Chimera SFA8+BN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8344)
  AUTHORS   Alcocer MJ, Murtagh GJ, Wilson PB, Progias P, Lin J, Archer DB
  TITLE     The major human structural IgE epitope of the Brazil nut allergen 
            Ber e 1: a chimaeric and protein microarray approach.
  JOURNAL   J Mol Biol. 2004 Oct 22;343(3):759-69. doi: 
            10.1016/j.jmb.2004.08.065.
  PUBMED    15465060
REFERENCE   2  (bases 1 to 8344)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.jmb.2004.08"; journalName: "J Mol Biol"; date: 
            "2004-10-22- 22"; volume: "343"; issue: "3"; pages: "759-69"
COMMENT     Sequence Label: pGJM015
FEATURES             Location/Qualifiers
     source          1..8344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..936
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             join(948..1004,1005..1202,1203..1214)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     terminator      1648..1894
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2307..4841)
                     /codon_start=1
                     /gene="Pichia pastoris HIS4"
                     /product="multifunctional enzyme, required for histidine 
                     biosynthesis"
                     /label=PpHIS4
                     /note="auxotrophic marker for Pichia pastoris"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGCKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     primer_bind     complement(4982..5001)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    5196..5944
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(5980..6002)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6088..6228
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6143..6162
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6243..6260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6414..7002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6494..6513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7173..7964,7965..8033))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7796..7815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8034..8138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8206..8224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gatctaacat ccaaagacga aaggttgaat gaaacctttt tgccatccga catccacagg
       61 tccattctca cacataagtg ccaaacgcaa caggagggga tacactagca gcagaccgtt
      121 gcaaacgcag gacctccact cctcttctcc tcaacaccca cttttgccat cgaaaaacca
      181 gcccagttat tgggcttgat tggagctcgc tcattccaat tccttctatt aggctactaa
      241 caccatgact ttattagcct gtctatcctg gcccccctgg cgaggttcat gtttgtttat
      301 ttccgaatgc aacaagctcc gcattacacc cgaacatcac tccagatgag ggctttctga
      361 gtgtggggtc aaatagtttc atgttcccca aatggcccaa aactgacagt ttaaacgctg
      421 tcttggaacc taatatgaca aaagcgtgat ctcatccaag atgaactaag tttggttcgt
      481 tgaaatgcta acggccagtt ggtcaaaaag aaacttccaa aagtcggcat accgtttgtc
      541 ttgtttggta ttgattgacg aatgctcaaa aataatctca ttaatgctta gcgcagtctc
      601 tctatcgctt ctgaaccccg gtgcacctgt gccgaaacgc aaatggggaa acacccgctt
      661 tttggatgat tatgcattgt ctccacattg tatgcttcca agattctggt gggaatactg
      721 ctgatagcct aacgttcatg atcaaaattt aactgttcta acccctactt gacagcaata
      781 tataaacaga aggaagctgc cctgtcttaa accttttttt ttatcatcat tattagctta
      841 ctttcataat tgcgactggt tccaattgac aagcttttga ttttaacgac ttttaacgac
      901 aacttgagaa gatcaaaaaa caactaatta ttcgaaggat ccaaacgatg agatttcctt
      961 caatttttac tgcagtttta ttcgcagcat cctccgcatt agctgctcca gtcaacacta
     1021 caacagaaga tgaaacggca caaattccgg ctgaagctgt catcggttac tcagatttag
     1081 aaggggattt cgatgttgct gttttgccat tttccaacag cacaaataac gggttattgt
     1141 ttataaatac tactattgcc agcattgctg ctaaagaaga aggggtatct ctcgagaaaa
     1201 gagaggctga agctccctat ggccgaggta gaactgaatc cggatgctat cagcagatgg
     1261 aggaggcgga gatgctcaac cactgcggga tgtacctcat gaaaaattta ggagaaagga
     1321 gccaagttag tcccagaatg agagaagagg atcacaagca actttgctgc atgcaactga
     1381 agaacctaga tgagaagtgc atgtgtccgg caatcatgat gatgttgaac gaaccaatgt
     1441 ggataaggat gcgagatcaa gtgatgagca tggctcataa cctgcccata gaatgcaacc
     1501 tcagtcccat gagatgcccc atgggtggct ccattgccgg gttctatgag aattccctag
     1561 ggcgccgcga attaattcgc cttagacatg actgttcctc agttcaagtt gggcacttac
     1621 gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat ttgcctgaga
     1681 gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata ggattttttt
     1741 tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg cagctgatga
     1801 atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct tggtatttcc
     1861 cactcctctt cagagtacag aagattaagt gagacgttcg tttgtgcaag cttatcgata
     1921 agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga
     1981 aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag
     2041 gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg
     2101 ccagtcacta tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg
     2161 ttctcggagc actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg
     2221 gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatc tatcgaatct
     2281 aaatgtaagt taaaatctct aaataattaa ataagtccca gtttctccat acgaacctta
     2341 acagcattgc ggtgagcatc tagaccttca acagcagcca gatccatcac tgcttggcca
     2401 atatgtttca gtccctcagg agttacgtct tgtgaagtga tgaacttctg gaaggttgca
     2461 gtgttaactc cgctgtattg acgggcatat ccgtacgttg gcaaagtgtg gttggtaccg
     2521 gaggagtaat ctccacaact ctctggagag taggcaccaa caaacacaga tccagcgtgt
     2581 tgtacttgat caacataaga agaagcattc tcgatttgca ggatcaagtg ttcaggagcg
     2641 tactgattgg acatttccaa agcctgctcg taggttgcaa ccgatagggt tgtagagtgt
     2701 gcaatacact tgcgtacaat ttcaaccctt ggcaactgca cagcttggtt gtgaacagca
     2761 tcttcaattc tggcaagctc cttgtctgtc atatcgacag ccaacagaat cacctgggaa
     2821 tcaataccat gttcagcttg agacagaagg tctgaggcaa cgaaatctgg atcagcgtat
     2881 ttatcagcaa taactagaac ttcagaaggc ccagcaggca tgtcaatact acacagggct
     2941 gatgtgtcat tttgaaccat catcttggca gcagtaacga actggtttcc tggaccaaat
     3001 attttgtcac acttaggaac agtttctgtt ccgtaagcca tagcagctac tgcctgggcg
     3061 cctcctgcta gcacgataca cttagcacca accttgtggg caacgtagat gacttctggg
     3121 gtaagggtac catccttctt aggtggagat gcaaaaacaa tttctttgca accagcaact
     3181 ttggcaggaa cacccagcat cagggaagtg gaaggcagaa ttgcggttcc accaggaata
     3241 tagaggccaa ctttctcaat aggtcttgca aaacgagagc agactacacc agggcaagtc
     3301 tcaacttgca acgtctccgt tagttgagct tcatggaatt tcctgacgtt atctatagag
     3361 agatcaatgg ctctcttaac gttatctggc aattgcataa gttcctctgg gaaaggagct
     3421 tctaacacag gtgtcttcaa agcgactcca tcaaacttgg cagttagttc taaaagggct
     3481 ttgtcaccat tttgacgaac attgtcgaca attggtttga ctaattccat aatctgttcc
     3541 gttttctgga taggacgacg aagggcatct tcaatttctt gtgaggaggc cttagaaacg
     3601 tcaattttgc acaattcaat acgaccttca gaagggactt ctttaggttt ggattcttct
     3661 ttaggttgtt ccttggtgta tcctggcttg gcatctcctt tccttctagt gacctttagg
     3721 gacttcatat ccaggtttct ctccacctcg tccaacgtca caccgtactt ggcacatcta
     3781 actaatgcaa aataaaataa gtcagcacat tcccaggcta tatcttcctt ggatttagct
     3841 tctgcaagtt catcagcttc ctccctaatt ttagcgttca acaaaacttc gtcgtcaaat
     3901 aaccgtttgg tataagaacc ttctggagca ttgctcttac gatcccacaa ggtggcttcc
     3961 atggctctaa gaccctttga ttggccaaaa caggaagtgc gttccaagtg acagaaacca
     4021 acacctgttt gttcaaccac aaatttcaag cagtctccat cacaatccaa ttcgataccc
     4081 agcaactttt gagttgctcc agatgtagca cctttatacc acaaaccgtg acgacgagat
     4141 tggtagactc cagtttgtgt ccttatagcc tccggaatag actttttgga cgagtacacc
     4201 aggcccaacg agtaattaga agagtcagcc accaaagtag tgaatagacc atcggggcgg
     4261 tcagtagtca aagacgccaa caaaatttca ctgacaggga actttttgac atcttcagaa
     4321 agttcgtatt cagtagtcaa ttgccgagca tcaataatgg ggattatacc agaagcaaca
     4381 gtggaagtca catctaccaa ctttgcggtc tcagaaaaag cataaacagt tctactaccg
     4441 ccattagtga aacttttcaa atcgcccagt ggagaagaaa aaggcacagc gatactagca
     4501 ttagcgggca aggatgcaac tttatcaacc agggtcctat agataaccct agcgcctggg
     4561 atcatccttt ggacaactct ttctgccaaa tctaggtcca aaatcacttc attgatacca
     4621 ttattgtaca acttgagcaa gttgtcgatc agctcctcaa attggtcctc tgtaacggat
     4681 gactcaactt gcacattaac ttgaagctca gtcgattgag tgaacttgat caggttgtgc
     4741 agctggtcag cagcataggg aaacacggct tttcctacca aactcaagga attatcaaac
     4801 tctgcaacac ttgcgtatgc aggtagcaag ggaaatgtca tacttgaagt cggacagtga
     4861 gtgtagtctt gagaaattct gaagccgtat ttttattatc agtgagtcag tcatcaggag
     4921 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     4981 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     5041 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
     5101 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
     5161 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag tatctatgat tggaagtatg
     5221 ggaatggtga tacccgcatt cttcagtgtc ttgaggtctc ctatcagatt atgcccaact
     5281 aaagcaaccg gaggaggaga tttcatggta aatttctctg acttttggtc atcagtagac
     5341 tcgaactgtg agactatctc ggttatgaca gcagaaatgt ccttcttgga gacagtaaat
     5401 gaagtcccac caataaagaa atccttgtta tcaggaacaa acttcttgtt tcgaactttt
     5461 tcggtgcctt gaactataaa atgtagagtg gatatgtcgg gtaggaatgg agcgggcaaa
     5521 tgcttacctt ctggaccttc aagaggtatg tagggtttgt agatactgat gccaacttca
     5581 gtgacaacgt tgctatttcg ttcaaaccat tccgaatcca gagaaatcaa agttgtttgt
     5641 ctactattga tccaagccag tgcggtcttg aaactgacaa tagtgtgctc gtgttttgag
     5701 gtcatctttg tatgaataaa tctagtcttt gatctaaata atcttgacga gccagacgat
     5761 aataccaatc taaactcttt aaacgttaaa ggacaagtat gtctgcctgt attaaacccc
     5821 aaatcagctc gtagtctgat cctcatcaac ttgaggggca ctatcttgtt ttagagaaat
     5881 ttgcggagat gcgatatcga gaaaaaggta cgctgatttt aaacgtgaaa tttatctcaa
     5941 gatctgctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6001 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6061 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     6121 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6181 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     6241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6841 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     7501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     7801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8221 aggccctttc gtcttcaaga attaattctc atgtttgaca gcttatcatc gataagctga
     8281 ctcatgttgg tattgtgaaa tagacgcaga tcgggaacac tgaaaaataa cagttattat
     8341 tcga
//
