LOCUS       sequence-251678-        8361 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 8361)
  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 8361)
  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: pGJM012
FEATURES             Location/Qualifiers
     source          1..8361
                     /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      1665..1911
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2324..4858)
                     /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(4999..5018)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    5213..5961
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(5997..6019)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6105..6245
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6160..6179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6260..6277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6431..7019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6511..6530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7190..7981,7982..8050))
                     /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     7813..7832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8051..8155)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8223..8241
                     /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 atgtgcgaag gcttaaggat gatgatgagg atgatgcaac
     1441 agaaggagat gcaaccccga ggggagcaga tgcgaaggat gatgaggctg gccgagaata
     1501 tcccttcccg ctgcaacctc agtcccatga gatgccccat gggtggctcc attgccgggt
     1561 tctgagaatt ccctagggcg gccgcgaatt aattcgcctt agacatgact gttcctcagt
     1621 tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga
     1681 atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata
     1741 tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc
     1801 tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg
     1861 tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag acgttcgttt
     1921 gtgcaagctt atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag
     1981 tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct
     2041 ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt
     2101 ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca
     2161 atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct
     2221 gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct
     2281 gtggatctat cgaatctaaa tgtaagttaa aatctctaaa taattaaata agtcccagtt
     2341 tctccatacg aaccttaaca gcattgcggt gagcatctag accttcaaca gcagccagat
     2401 ccatcactgc ttggccaata tgtttcagtc cctcaggagt tacgtcttgt gaagtgatga
     2461 acttctggaa ggttgcagtg ttaactccgc tgtattgacg ggcatatccg tacgttggca
     2521 aagtgtggtt ggtaccggag gagtaatctc cacaactctc tggagagtag gcaccaacaa
     2581 acacagatcc agcgtgttgt acttgatcaa cataagaaga agcattctcg atttgcagga
     2641 tcaagtgttc aggagcgtac tgattggaca tttccaaagc ctgctcgtag gttgcaaccg
     2701 atagggttgt agagtgtgca atacacttgc gtacaatttc aacccttggc aactgcacag
     2761 cttggttgtg aacagcatct tcaattctgg caagctcctt gtctgtcata tcgacagcca
     2821 acagaatcac ctgggaatca ataccatgtt cagcttgaga cagaaggtct gaggcaacga
     2881 aatctggatc agcgtattta tcagcaataa ctagaacttc agaaggccca gcaggcatgt
     2941 caatactaca cagggctgat gtgtcatttt gaaccatcat cttggcagca gtaacgaact
     3001 ggtttcctgg accaaatatt ttgtcacact taggaacagt ttctgttccg taagccatag
     3061 cagctactgc ctgggcgcct cctgctagca cgatacactt agcaccaacc ttgtgggcaa
     3121 cgtagatgac ttctggggta agggtaccat ccttcttagg tggagatgca aaaacaattt
     3181 ctttgcaacc agcaactttg gcaggaacac ccagcatcag ggaagtggaa ggcagaattg
     3241 cggttccacc aggaatatag aggccaactt tctcaatagg tcttgcaaaa cgagagcaga
     3301 ctacaccagg gcaagtctca acttgcaacg tctccgttag ttgagcttca tggaatttcc
     3361 tgacgttatc tatagagaga tcaatggctc tcttaacgtt atctggcaat tgcataagtt
     3421 cctctgggaa aggagcttct aacacaggtg tcttcaaagc gactccatca aacttggcag
     3481 ttagttctaa aagggctttg tcaccatttt gacgaacatt gtcgacaatt ggtttgacta
     3541 attccataat ctgttccgtt ttctggatag gacgacgaag ggcatcttca atttcttgtg
     3601 aggaggcctt agaaacgtca attttgcaca attcaatacg accttcagaa gggacttctt
     3661 taggtttgga ttcttcttta ggttgttcct tggtgtatcc tggcttggca tctcctttcc
     3721 ttctagtgac ctttagggac ttcatatcca ggtttctctc cacctcgtcc aacgtcacac
     3781 cgtacttggc acatctaact aatgcaaaat aaaataagtc agcacattcc caggctatat
     3841 cttccttgga tttagcttct gcaagttcat cagcttcctc cctaatttta gcgttcaaca
     3901 aaacttcgtc gtcaaataac cgtttggtat aagaaccttc tggagcattg ctcttacgat
     3961 cccacaaggt ggcttccatg gctctaagac cctttgattg gccaaaacag gaagtgcgtt
     4021 ccaagtgaca gaaaccaaca cctgtttgtt caaccacaaa tttcaagcag tctccatcac
     4081 aatccaattc gatacccagc aacttttgag ttgctccaga tgtagcacct ttataccaca
     4141 aaccgtgacg acgagattgg tagactccag tttgtgtcct tatagcctcc ggaatagact
     4201 ttttggacga gtacaccagg cccaacgagt aattagaaga gtcagccacc aaagtagtga
     4261 atagaccatc ggggcggtca gtagtcaaag acgccaacaa aatttcactg acagggaact
     4321 ttttgacatc ttcagaaagt tcgtattcag tagtcaattg ccgagcatca ataatgggga
     4381 ttataccaga agcaacagtg gaagtcacat ctaccaactt tgcggtctca gaaaaagcat
     4441 aaacagttct actaccgcca ttagtgaaac ttttcaaatc gcccagtgga gaagaaaaag
     4501 gcacagcgat actagcatta gcgggcaagg atgcaacttt atcaaccagg gtcctataga
     4561 taaccctagc gcctgggatc atcctttgga caactctttc tgccaaatct aggtccaaaa
     4621 tcacttcatt gataccatta ttgtacaact tgagcaagtt gtcgatcagc tcctcaaatt
     4681 ggtcctctgt aacggatgac tcaacttgca cattaacttg aagctcagtc gattgagtga
     4741 acttgatcag gttgtgcagc tggtcagcag catagggaaa cacggctttt cctaccaaac
     4801 tcaaggaatt atcaaactct gcaacacttg cgtatgcagg tagcaaggga aatgtcatac
     4861 ttgaagtcgg acagtgagtg tagtcttgag aaattctgaa gccgtatttt tattatcagt
     4921 gagtcagtca tcaggagatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     4981 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     5041 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     5101 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     5161 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagtat
     5221 ctatgattgg aagtatggga atggtgatac ccgcattctt cagtgtcttg aggtctccta
     5281 tcagattatg cccaactaaa gcaaccggag gaggagattt catggtaaat ttctctgact
     5341 tttggtcatc agtagactcg aactgtgaga ctatctcggt tatgacagca gaaatgtcct
     5401 tcttggagac agtaaatgaa gtcccaccaa taaagaaatc cttgttatca ggaacaaact
     5461 tcttgtttcg aactttttcg gtgccttgaa ctataaaatg tagagtggat atgtcgggta
     5521 ggaatggagc gggcaaatgc ttaccttctg gaccttcaag aggtatgtag ggtttgtaga
     5581 tactgatgcc aacttcagtg acaacgttgc tatttcgttc aaaccattcc gaatccagag
     5641 aaatcaaagt tgtttgtcta ctattgatcc aagccagtgc ggtcttgaaa ctgacaatag
     5701 tgtgctcgtg ttttgaggtc atctttgtat gaataaatct agtctttgat ctaaataatc
     5761 ttgacgagcc agacgataat accaatctaa actctttaaa cgttaaagga caagtatgtc
     5821 tgcctgtatt aaaccccaaa tcagctcgta gtctgatcct catcaacttg aggggcacta
     5881 tcttgtttta gagaaatttg cggagatgcg atatcgagaa aaaggtacgc tgattttaaa
     5941 cgtgaaattt atctcaagat ctgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6001 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6061 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     6121 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     6181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6241 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6841 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7321 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7501 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7561 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7621 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7681 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7741 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7801 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7861 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7921 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7981 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8041 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8101 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8161 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     8221 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt aattctcatg tttgacagct
     8281 tatcatcgat aagctgactc atgttggtat tgtgaaatag acgcagatcg ggaacactga
     8341 aaaataacag ttattattcg a
//
