LOCUS       sequence-251719-        8332 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 8332)
  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 8332)
  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: pGJM013
FEATURES             Location/Qualifiers
     source          1..8332
                     /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      1636..1882
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2295..4829)
                     /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(4970..4989)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    5184..5932
                     /label=AOX1 3' fragment
                     /note="region downstream of Pichia pastoris AOX1 gene"
     primer_bind     complement(5968..5990)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6076..6216
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6131..6150
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6231..6248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6402..6990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6482..6501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7161..7952,7953..8021))
                     /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     7784..7803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8022..8126)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8194..8212
                     /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 cactgccgga tggatatgag acagcagatg gaggagagca
     1321 cgtaccagac catgcccagg cggggaatgg agccgcatat gagcgagtgc tgcatgcaac
     1381 tgaagaacct agatgagaag tgcatgtgtc cggcaatcat gatgatgttg aacgaaccaa
     1441 tgtggataag gatgcgagat caagtgatga gcatggctca taacctgccc atagaatgca
     1501 atctcatgtc ccaaccatgc caaatgtaat aagggcgaat tccctagggc ggccgcgaat
     1561 taattcgcct tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt
     1621 cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc
     1681 atttttgata cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt
     1741 tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt
     1801 aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca
     1861 gagtacagaa gattaagtga gacgttcgtt tgtgcaagct tatcgataag ctttaatgcg
     1921 gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg
     1981 cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc
     2041 cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg
     2101 gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac
     2161 tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg
     2221 actacgcgat catggcgacc acacccgtcc tgtggatcta tcgaatctaa atgtaagtta
     2281 aaatctctaa ataattaaat aagtcccagt ttctccatac gaaccttaac agcattgcgg
     2341 tgagcatcta gaccttcaac agcagccaga tccatcactg cttggccaat atgtttcagt
     2401 ccctcaggag ttacgtcttg tgaagtgatg aacttctgga aggttgcagt gttaactccg
     2461 ctgtattgac gggcatatcc gtacgttggc aaagtgtggt tggtaccgga ggagtaatct
     2521 ccacaactct ctggagagta ggcaccaaca aacacagatc cagcgtgttg tacttgatca
     2581 acataagaag aagcattctc gatttgcagg atcaagtgtt caggagcgta ctgattggac
     2641 atttccaaag cctgctcgta ggttgcaacc gatagggttg tagagtgtgc aatacacttg
     2701 cgtacaattt caacccttgg caactgcaca gcttggttgt gaacagcatc ttcaattctg
     2761 gcaagctcct tgtctgtcat atcgacagcc aacagaatca cctgggaatc aataccatgt
     2821 tcagcttgag acagaaggtc tgaggcaacg aaatctggat cagcgtattt atcagcaata
     2881 actagaactt cagaaggccc agcaggcatg tcaatactac acagggctga tgtgtcattt
     2941 tgaaccatca tcttggcagc agtaacgaac tggtttcctg gaccaaatat tttgtcacac
     3001 ttaggaacag tttctgttcc gtaagccata gcagctactg cctgggcgcc tcctgctagc
     3061 acgatacact tagcaccaac cttgtgggca acgtagatga cttctggggt aagggtacca
     3121 tccttcttag gtggagatgc aaaaacaatt tctttgcaac cagcaacttt ggcaggaaca
     3181 cccagcatca gggaagtgga aggcagaatt gcggttccac caggaatata gaggccaact
     3241 ttctcaatag gtcttgcaaa acgagagcag actacaccag ggcaagtctc aacttgcaac
     3301 gtctccgtta gttgagcttc atggaatttc ctgacgttat ctatagagag atcaatggct
     3361 ctcttaacgt tatctggcaa ttgcataagt tcctctggga aaggagcttc taacacaggt
     3421 gtcttcaaag cgactccatc aaacttggca gttagttcta aaagggcttt gtcaccattt
     3481 tgacgaacat tgtcgacaat tggtttgact aattccataa tctgttccgt tttctggata
     3541 ggacgacgaa gggcatcttc aatttcttgt gaggaggcct tagaaacgtc aattttgcac
     3601 aattcaatac gaccttcaga agggacttct ttaggtttgg attcttcttt aggttgttcc
     3661 ttggtgtatc ctggcttggc atctcctttc cttctagtga cctttaggga cttcatatcc
     3721 aggtttctct ccacctcgtc caacgtcaca ccgtacttgg cacatctaac taatgcaaaa
     3781 taaaataagt cagcacattc ccaggctata tcttccttgg atttagcttc tgcaagttca
     3841 tcagcttcct ccctaatttt agcgttcaac aaaacttcgt cgtcaaataa ccgtttggta
     3901 taagaacctt ctggagcatt gctcttacga tcccacaagg tggcttccat ggctctaaga
     3961 ccctttgatt ggccaaaaca ggaagtgcgt tccaagtgac agaaaccaac acctgtttgt
     4021 tcaaccacaa atttcaagca gtctccatca caatccaatt cgatacccag caacttttga
     4081 gttgctccag atgtagcacc tttataccac aaaccgtgac gacgagattg gtagactcca
     4141 gtttgtgtcc ttatagcctc cggaatagac tttttggacg agtacaccag gcccaacgag
     4201 taattagaag agtcagccac caaagtagtg aatagaccat cggggcggtc agtagtcaaa
     4261 gacgccaaca aaatttcact gacagggaac tttttgacat cttcagaaag ttcgtattca
     4321 gtagtcaatt gccgagcatc aataatgggg attataccag aagcaacagt ggaagtcaca
     4381 tctaccaact ttgcggtctc agaaaaagca taaacagttc tactaccgcc attagtgaaa
     4441 cttttcaaat cgcccagtgg agaagaaaaa ggcacagcga tactagcatt agcgggcaag
     4501 gatgcaactt tatcaaccag ggtcctatag ataaccctag cgcctgggat catcctttgg
     4561 acaactcttt ctgccaaatc taggtccaaa atcacttcat tgataccatt attgtacaac
     4621 ttgagcaagt tgtcgatcag ctcctcaaat tggtcctctg taacggatga ctcaacttgc
     4681 acattaactt gaagctcagt cgattgagtg aacttgatca ggttgtgcag ctggtcagca
     4741 gcatagggaa acacggcttt tcctaccaaa ctcaaggaat tatcaaactc tgcaacactt
     4801 gcgtatgcag gtagcaaggg aaatgtcata cttgaagtcg gacagtgagt gtagtcttga
     4861 gaaattctga agccgtattt ttattatcag tgagtcagtc atcaggagat cctctacgcc
     4921 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     4981 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     5041 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     5101 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     5161 caggagtcgc ataagggaga gcgtcgagta tctatgattg gaagtatggg aatggtgata
     5221 cccgcattct tcagtgtctt gaggtctcct atcagattat gcccaactaa agcaaccgga
     5281 ggaggagatt tcatggtaaa tttctctgac ttttggtcat cagtagactc gaactgtgag
     5341 actatctcgg ttatgacagc agaaatgtcc ttcttggaga cagtaaatga agtcccacca
     5401 ataaagaaat ccttgttatc aggaacaaac ttcttgtttc gaactttttc ggtgccttga
     5461 actataaaat gtagagtgga tatgtcgggt aggaatggag cgggcaaatg cttaccttct
     5521 ggaccttcaa gaggtatgta gggtttgtag atactgatgc caacttcagt gacaacgttg
     5581 ctatttcgtt caaaccattc cgaatccaga gaaatcaaag ttgtttgtct actattgatc
     5641 caagccagtg cggtcttgaa actgacaata gtgtgctcgt gttttgaggt catctttgta
     5701 tgaataaatc tagtctttga tctaaataat cttgacgagc cagacgataa taccaatcta
     5761 aactctttaa acgttaaagg acaagtatgt ctgcctgtat taaaccccaa atcagctcgt
     5821 agtctgatcc tcatcaactt gaggggcact atcttgtttt agagaaattt gcggagatgc
     5881 gatatcgaga aaaaggtacg ctgattttaa acgtgaaatt tatctcaaga tctgctgcct
     5941 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6001 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6061 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
     6121 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6181 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     6241 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6301 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6361 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6421 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6481 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6541 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6601 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6661 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6721 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6781 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6841 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6901 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6961 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7021 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7081 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7141 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7201 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7261 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7321 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7381 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7441 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     7501 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7561 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7621 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7681 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7741 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     7801 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7861 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7921 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7981 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8041 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8101 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8161 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8221 cttcaagaat taattctcat gtttgacagc ttatcatcga taagctgact catgttggta
     8281 ttgtgaaata gacgcagatc gggaacactg aaaaataaca gttattattc ga
//
