LOCUS       sequence-174104-        3642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pichia pastoris expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3642)
  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 3642)
  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: pGHYB
FEATURES             Location/Qualifiers
     source          1..3642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      63..310
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(79..97)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(385..973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(465..484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1064..1540
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             1619..1648
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1664..1681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1761..2007
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     gene            2066..3641
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2066..2409
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2410..3438
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             2410..3438
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3444..3641
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 tgtcgattcg atactaacgc cgccatccag tgtcgaaaac gagctcgaat tcatcgatga
       61 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
      121 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
      181 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
      241 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
      301 tttaatttgc aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1021 aaactcacgt taagggattt tggtcatgca tgagatcaga tcttttttgt agaaatgtct
     1081 tggtgtcctc gtccaatcag gtagccatct ctgaaatatc tggctccgtt gcaactccga
     1141 acgacctgct ggcaacgtaa aattctccgg ggtaaaactt aaatgtggag taatggaacc
     1201 agaaacgtct cttcccttct ctctccttcc accgcccgtt accgtcccta ggaaatttta
     1261 ctctgctgga gagcttcttc tacggccccc ttgcagcaat gctcttccca gcattacgtt
     1321 gcgggtaaaa cggaggtcgt gtacccgacc tagcagccca gggatggaaa agtcccggcc
     1381 gtcgctggca ataatagcgg gcggacgcat gtcatgagat tattggaaac caccagaatc
     1441 gaatataaaa ggcgaacacc tttcccaatt ttggtttctc ctgacccaaa gactttaaat
     1501 ttaatttatt tgtccctatt tcaatcaatt gaacaactat ttcgaaacga ggaattcacg
     1561 tggcccagcc ggccgtctcg gatcggtacc tcgagccgcg gcggccgcca gctttctaga
     1621 acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca
     1681 ttgagtttgt agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
     1741 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     1801 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     1861 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     1921 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     1981 cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatctgttt agcttgcctc
     2041 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
     2101 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
     2161 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
     2221 cggctcctcg ctgcggacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
     2281 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     2341 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     2401 taaacaacca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     2461 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     2521 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     2581 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     2641 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     2701 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     2761 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     2821 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     2881 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     2941 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     3001 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     3061 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     3121 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     3181 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     3241 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     3301 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     3361 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     3421 ccgagggcaa aggaataatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     3481 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     3541 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     3601 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gc
//
