LOCUS       sequence-251563-        8349 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 8349)
  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 8349)
  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: pGJM010
FEATURES             Location/Qualifiers
     source          1..8349
                     /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..1201,1202..1213)
                     /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
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKERLK"
     terminator      1653..1899
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2312..4846)
                     /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(4987..5006)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    5201..5949
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(5985..6007)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6093..6233
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6148..6167
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6248..6265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6419..7007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6499..6518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7178..7969,7970..8038))
                     /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     7801..7820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8039..8143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8211..8229
                     /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 ctcgagaaag
     1201 agaggctgaa gctccctatg gccgaggtag aactgaatcc ggatgctatc agcagatgga
     1261 ggaggcggag atgctcaacc actgcgggat gtacctcatg aaaaatttag gagaaaggag
     1321 ccaagttagt cccagaatga gagaagagga tcacaagcaa ctttgctgca tgcaactgaa
     1381 gaacctagat gagaagtgca tgtgcgaagg cttaaggatg atgatgagga tgatgcaaca
     1441 gaagttgaac gaaccaatgt ggataaggat gcgagatcaa gtgatgagca tggctcataa
     1501 cctgcccata gaatgcaatc tcatgtccca accatgccaa atgtaataag ggcgaattcc
     1561 ctagggcggc cgcgaattaa ttcgccttag acatgactgt tcctcagttc aagttgggca
     1621 cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc
     1681 tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt
     1741 ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct
     1801 gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta
     1861 tttcccactc ctcttcagag tacagaagat taagtgagac gttcgtttgt gcaagcttat
     1921 cgataagctt taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg
     1981 tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg
     2041 cataggcttg gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag
     2101 catcgccagt cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc
     2161 acccgttctc ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct
     2221 acttggagcc actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggatctatcg
     2281 aatctaaatg taagttaaaa tctctaaata attaaataag tcccagtttc tccatacgaa
     2341 ccttaacagc attgcggtga gcatctagac cttcaacagc agccagatcc atcactgctt
     2401 ggccaatatg tttcagtccc tcaggagtta cgtcttgtga agtgatgaac ttctggaagg
     2461 ttgcagtgtt aactccgctg tattgacggg catatccgta cgttggcaaa gtgtggttgg
     2521 taccggagga gtaatctcca caactctctg gagagtaggc accaacaaac acagatccag
     2581 cgtgttgtac ttgatcaaca taagaagaag cattctcgat ttgcaggatc aagtgttcag
     2641 gagcgtactg attggacatt tccaaagcct gctcgtaggt tgcaaccgat agggttgtag
     2701 agtgtgcaat acacttgcgt acaatttcaa cccttggcaa ctgcacagct tggttgtgaa
     2761 cagcatcttc aattctggca agctccttgt ctgtcatatc gacagccaac agaatcacct
     2821 gggaatcaat accatgttca gcttgagaca gaaggtctga ggcaacgaaa tctggatcag
     2881 cgtatttatc agcaataact agaacttcag aaggcccagc aggcatgtca atactacaca
     2941 gggctgatgt gtcattttga accatcatct tggcagcagt aacgaactgg tttcctggac
     3001 caaatatttt gtcacactta ggaacagttt ctgttccgta agccatagca gctactgcct
     3061 gggcgcctcc tgctagcacg atacacttag caccaacctt gtgggcaacg tagatgactt
     3121 ctggggtaag ggtaccatcc ttcttaggtg gagatgcaaa aacaatttct ttgcaaccag
     3181 caactttggc aggaacaccc agcatcaggg aagtggaagg cagaattgcg gttccaccag
     3241 gaatatagag gccaactttc tcaataggtc ttgcaaaacg agagcagact acaccagggc
     3301 aagtctcaac ttgcaacgtc tccgttagtt gagcttcatg gaatttcctg acgttatcta
     3361 tagagagatc aatggctctc ttaacgttat ctggcaattg cataagttcc tctgggaaag
     3421 gagcttctaa cacaggtgtc ttcaaagcga ctccatcaaa cttggcagtt agttctaaaa
     3481 gggctttgtc accattttga cgaacattgt cgacaattgg tttgactaat tccataatct
     3541 gttccgtttt ctggatagga cgacgaaggg catcttcaat ttcttgtgag gaggccttag
     3601 aaacgtcaat tttgcacaat tcaatacgac cttcagaagg gacttcttta ggtttggatt
     3661 cttctttagg ttgttccttg gtgtatcctg gcttggcatc tcctttcctt ctagtgacct
     3721 ttagggactt catatccagg tttctctcca cctcgtccaa cgtcacaccg tacttggcac
     3781 atctaactaa tgcaaaataa aataagtcag cacattccca ggctatatct tccttggatt
     3841 tagcttctgc aagttcatca gcttcctccc taattttagc gttcaacaaa acttcgtcgt
     3901 caaataaccg tttggtataa gaaccttctg gagcattgct cttacgatcc cacaaggtgg
     3961 cttccatggc tctaagaccc tttgattggc caaaacagga agtgcgttcc aagtgacaga
     4021 aaccaacacc tgtttgttca accacaaatt tcaagcagtc tccatcacaa tccaattcga
     4081 tacccagcaa cttttgagtt gctccagatg tagcaccttt ataccacaaa ccgtgacgac
     4141 gagattggta gactccagtt tgtgtcctta tagcctccgg aatagacttt ttggacgagt
     4201 acaccaggcc caacgagtaa ttagaagagt cagccaccaa agtagtgaat agaccatcgg
     4261 ggcggtcagt agtcaaagac gccaacaaaa tttcactgac agggaacttt ttgacatctt
     4321 cagaaagttc gtattcagta gtcaattgcc gagcatcaat aatggggatt ataccagaag
     4381 caacagtgga agtcacatct accaactttg cggtctcaga aaaagcataa acagttctac
     4441 taccgccatt agtgaaactt ttcaaatcgc ccagtggaga agaaaaaggc acagcgatac
     4501 tagcattagc gggcaaggat gcaactttat caaccagggt cctatagata accctagcgc
     4561 ctgggatcat cctttggaca actctttctg ccaaatctag gtccaaaatc acttcattga
     4621 taccattatt gtacaacttg agcaagttgt cgatcagctc ctcaaattgg tcctctgtaa
     4681 cggatgactc aacttgcaca ttaacttgaa gctcagtcga ttgagtgaac ttgatcaggt
     4741 tgtgcagctg gtcagcagca tagggaaaca cggcttttcc taccaaactc aaggaattat
     4801 caaactctgc aacacttgcg tatgcaggta gcaagggaaa tgtcatactt gaagtcggac
     4861 agtgagtgta gtcttgagaa attctgaagc cgtattttta ttatcagtga gtcagtcatc
     4921 aggagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     4981 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     5041 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     5101 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     5161 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagtatct atgattggaa
     5221 gtatgggaat ggtgataccc gcattcttca gtgtcttgag gtctcctatc agattatgcc
     5281 caactaaagc aaccggagga ggagatttca tggtaaattt ctctgacttt tggtcatcag
     5341 tagactcgaa ctgtgagact atctcggtta tgacagcaga aatgtccttc ttggagacag
     5401 taaatgaagt cccaccaata aagaaatcct tgttatcagg aacaaacttc ttgtttcgaa
     5461 ctttttcggt gccttgaact ataaaatgta gagtggatat gtcgggtagg aatggagcgg
     5521 gcaaatgctt accttctgga ccttcaagag gtatgtaggg tttgtagata ctgatgccaa
     5581 cttcagtgac aacgttgcta tttcgttcaa accattccga atccagagaa atcaaagttg
     5641 tttgtctact attgatccaa gccagtgcgg tcttgaaact gacaatagtg tgctcgtgtt
     5701 ttgaggtcat ctttgtatga ataaatctag tctttgatct aaataatctt gacgagccag
     5761 acgataatac caatctaaac tctttaaacg ttaaaggaca agtatgtctg cctgtattaa
     5821 accccaaatc agctcgtagt ctgatcctca tcaacttgag gggcactatc ttgttttaga
     5881 gaaatttgcg gagatgcgat atcgagaaaa aggtacgctg attttaaacg tgaaatttat
     5941 ctcaagatct gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6001 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6061 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     6121 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6181 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     6241 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6841 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7081 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     7501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     7801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8161 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     8221 tcacgaggcc ctttcgtctt caagaattaa ttctcatgtt tgacagctta tcatcgataa
     8281 gctgactcat gttggtattg tgaaatagac gcagatcggg aacactgaaa aataacagtt
     8341 attattcga
//
