LOCUS       sequence-174102-        3506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression in Pichia pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3506)
  AUTHORS   Yang J, Nie L, Chen B, Liu Y, Kong Y, Wang H, Diao L
  TITLE     Hygromycin-resistance vectors for gene expression in Pichia 
            pastoris.
  JOURNAL   Yeast. 2014 Apr;31(4):115-25.
  PUBMED    24822243
REFERENCE   2  (bases 1 to 3506)
  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"; journalName: "Yeast."; 
            date: "2014-04"; volume: "31(4)"; pages: "115-25"
COMMENT     Sequence Label: pGKB
FEATURES             Location/Qualifiers
     source          1..3506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        58..534
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             613..642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             658..675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      755..1001
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1016..1429
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1436..1483
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1502..2311
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1569..1588)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2317..2514
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     terminator      2563..2810
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2579..2597)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2885..3473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2965..2984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcatgagat cagatctttt
       61 ttgtagaaat gtcttggtgt cctcgtccaa tcaggtagcc atctctgaaa tatctggctc
      121 cgttgcaact ccgaacgacc tgctggcaac gtaaaattct ccggggtaaa acttaaatgt
      181 ggagtaatgg aaccagaaac gtctcttccc ttctctctcc ttccaccgcc cgttaccgtc
      241 cctaggaaat tttactctgc tggagagctt cttctacggc ccccttgcag caatgctctt
      301 cccagcatta cgttgcgggt aaaacggagg tcgtgtaccc gacctagcag cccagggatg
      361 gaaaagtccc ggccgtcgct ggcaataata gcgggcggac gcatgtcatg agattattgg
      421 aaaccaccag aatcgaatat aaaaggcgaa cacctttccc aattttggtt tctcctgacc
      481 caaagacttt aaatttaatt tatttgtccc tatttcaatc aattgaacaa ctatttcgaa
      541 acgaggaatt cacgtggccc agccggccgt ctcggatcgg tacctcgagc cgcggcggcc
      601 gccagctttc tagaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
      661 catcatcatc atcattgagt ttgtagcctt agacatgact gttcctcagt tcaagttggg
      721 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg
      781 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
      841 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
      901 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
      961 tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatcc
     1021 cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     1081 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     1141 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     1201 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     1261 tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
     1321 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
     1381 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagg gcggtgttga
     1441 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     1501 catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga
     1561 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1621 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1681 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1741 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1801 cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1861 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1921 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1981 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2041 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2101 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2161 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2221 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2281 gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt
     2341 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     2401 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     2461 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     2521 attcgatact aacgccgcca tccagtgtcg aaaacgatat catcatgtaa ttagttatgt
     2581 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     2641 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     2701 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     2761 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctggaga
     2821 ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3481 ctttgatctt ttctacgggg tctgac
//
