LOCUS       sequence-251360-        8329 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 8329)
  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 8329)
  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: pGJM09
FEATURES             Location/Qualifiers
     source          1..8329
                     /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      1633..1879
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2292..4826)
                     /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(4967..4986)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    5181..5929
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(5965..5987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6073..6213
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6128..6147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6228..6245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6399..6987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6479..6498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7158..7949,7950..8018))
                     /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     7781..7800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8019..8123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8191..8209
                     /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 gagcagctgg
     1381 aggggatgga cgagagctgc atgtgtccgg caatcatgat gatgttgaac gaaccaatgt
     1441 ggataaggat gcgagatcaa gtgatgagca tggctcataa cctgcccata gaatgcaatc
     1501 tcatgtccca accatgccaa atgtaataag ggcgaattcc ctagggcggc cgcgaattaa
     1561 ttcgccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt
     1621 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt
     1681 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct
     1741 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg
     1801 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
     1861 tacagaagat taagtgagac gttcgtttgt gcaagcttat cgataagctt taatgcggta
     1921 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     1981 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     2041 tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg
     2101 tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt
     2161 ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact
     2221 acgcgatcat ggcgaccaca cccgtcctgt ggatctatcg aatctaaatg taagttaaaa
     2281 tctctaaata attaaataag tcccagtttc tccatacgaa ccttaacagc attgcggtga
     2341 gcatctagac cttcaacagc agccagatcc atcactgctt ggccaatatg tttcagtccc
     2401 tcaggagtta cgtcttgtga agtgatgaac ttctggaagg ttgcagtgtt aactccgctg
     2461 tattgacggg catatccgta cgttggcaaa gtgtggttgg taccggagga gtaatctcca
     2521 caactctctg gagagtaggc accaacaaac acagatccag cgtgttgtac ttgatcaaca
     2581 taagaagaag cattctcgat ttgcaggatc aagtgttcag gagcgtactg attggacatt
     2641 tccaaagcct gctcgtaggt tgcaaccgat agggttgtag agtgtgcaat acacttgcgt
     2701 acaatttcaa cccttggcaa ctgcacagct tggttgtgaa cagcatcttc aattctggca
     2761 agctccttgt ctgtcatatc gacagccaac agaatcacct gggaatcaat accatgttca
     2821 gcttgagaca gaaggtctga ggcaacgaaa tctggatcag cgtatttatc agcaataact
     2881 agaacttcag aaggcccagc aggcatgtca atactacaca gggctgatgt gtcattttga
     2941 accatcatct tggcagcagt aacgaactgg tttcctggac caaatatttt gtcacactta
     3001 ggaacagttt ctgttccgta agccatagca gctactgcct gggcgcctcc tgctagcacg
     3061 atacacttag caccaacctt gtgggcaacg tagatgactt ctggggtaag ggtaccatcc
     3121 ttcttaggtg gagatgcaaa aacaatttct ttgcaaccag caactttggc aggaacaccc
     3181 agcatcaggg aagtggaagg cagaattgcg gttccaccag gaatatagag gccaactttc
     3241 tcaataggtc ttgcaaaacg agagcagact acaccagggc aagtctcaac ttgcaacgtc
     3301 tccgttagtt gagcttcatg gaatttcctg acgttatcta tagagagatc aatggctctc
     3361 ttaacgttat ctggcaattg cataagttcc tctgggaaag gagcttctaa cacaggtgtc
     3421 ttcaaagcga ctccatcaaa cttggcagtt agttctaaaa gggctttgtc accattttga
     3481 cgaacattgt cgacaattgg tttgactaat tccataatct gttccgtttt ctggatagga
     3541 cgacgaaggg catcttcaat ttcttgtgag gaggccttag aaacgtcaat tttgcacaat
     3601 tcaatacgac cttcagaagg gacttcttta ggtttggatt cttctttagg ttgttccttg
     3661 gtgtatcctg gcttggcatc tcctttcctt ctagtgacct ttagggactt catatccagg
     3721 tttctctcca cctcgtccaa cgtcacaccg tacttggcac atctaactaa tgcaaaataa
     3781 aataagtcag cacattccca ggctatatct tccttggatt tagcttctgc aagttcatca
     3841 gcttcctccc taattttagc gttcaacaaa acttcgtcgt caaataaccg tttggtataa
     3901 gaaccttctg gagcattgct cttacgatcc cacaaggtgg cttccatggc tctaagaccc
     3961 tttgattggc caaaacagga agtgcgttcc aagtgacaga aaccaacacc tgtttgttca
     4021 accacaaatt tcaagcagtc tccatcacaa tccaattcga tacccagcaa cttttgagtt
     4081 gctccagatg tagcaccttt ataccacaaa ccgtgacgac gagattggta gactccagtt
     4141 tgtgtcctta tagcctccgg aatagacttt ttggacgagt acaccaggcc caacgagtaa
     4201 ttagaagagt cagccaccaa agtagtgaat agaccatcgg ggcggtcagt agtcaaagac
     4261 gccaacaaaa tttcactgac agggaacttt ttgacatctt cagaaagttc gtattcagta
     4321 gtcaattgcc gagcatcaat aatggggatt ataccagaag caacagtgga agtcacatct
     4381 accaactttg cggtctcaga aaaagcataa acagttctac taccgccatt agtgaaactt
     4441 ttcaaatcgc ccagtggaga agaaaaaggc acagcgatac tagcattagc gggcaaggat
     4501 gcaactttat caaccagggt cctatagata accctagcgc ctgggatcat cctttggaca
     4561 actctttctg ccaaatctag gtccaaaatc acttcattga taccattatt gtacaacttg
     4621 agcaagttgt cgatcagctc ctcaaattgg tcctctgtaa cggatgactc aacttgcaca
     4681 ttaacttgaa gctcagtcga ttgagtgaac ttgatcaggt tgtgcagctg gtcagcagca
     4741 tagggaaaca cggcttttcc taccaaactc aaggaattat caaactctgc aacacttgcg
     4801 tatgcaggta gcaagggaaa tgtcatactt gaagtcggac agtgagtgta gtcttgagaa
     4861 attctgaagc cgtattttta ttatcagtga gtcagtcatc aggagatcct ctacgccgga
     4921 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     4981 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     5041 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     5101 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     5161 gagtcgcata agggagagcg tcgagtatct atgattggaa gtatgggaat ggtgataccc
     5221 gcattcttca gtgtcttgag gtctcctatc agattatgcc caactaaagc aaccggagga
     5281 ggagatttca tggtaaattt ctctgacttt tggtcatcag tagactcgaa ctgtgagact
     5341 atctcggtta tgacagcaga aatgtccttc ttggagacag taaatgaagt cccaccaata
     5401 aagaaatcct tgttatcagg aacaaacttc ttgtttcgaa ctttttcggt gccttgaact
     5461 ataaaatgta gagtggatat gtcgggtagg aatggagcgg gcaaatgctt accttctgga
     5521 ccttcaagag gtatgtaggg tttgtagata ctgatgccaa cttcagtgac aacgttgcta
     5581 tttcgttcaa accattccga atccagagaa atcaaagttg tttgtctact attgatccaa
     5641 gccagtgcgg tcttgaaact gacaatagtg tgctcgtgtt ttgaggtcat ctttgtatga
     5701 ataaatctag tctttgatct aaataatctt gacgagccag acgataatac caatctaaac
     5761 tctttaaacg ttaaaggaca agtatgtctg cctgtattaa accccaaatc agctcgtagt
     5821 ctgatcctca tcaacttgag gggcactatc ttgttttaga gaaatttgcg gagatgcgat
     5881 atcgagaaaa aggtacgctg attttaaacg tgaaatttat ctcaagatct gctgcctcgc
     5941 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6001 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6061 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     6121 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     6181 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     6241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7441 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     7501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7741 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     7801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8161 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     8221 caagaattaa ttctcatgtt tgacagctta tcatcgataa gctgactcat gttggtattg
     8281 tgaaatagac gcagatcggg aacactgaaa aataacagtt attattcga
//
