LOCUS       sequence-267729-        4253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant yeast STE2 coding sequence in vector for inducible expression
            in the methanogenic yeast Pichia pastoris..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4253)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 4253)
  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"
COMMENT     Sequence Label: pPICZac-STE2-1D4
FEATURES             Location/Qualifiers
     source          1..4253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..939
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1934..1963
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1979..1996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2075..2321
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2336..2749
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2756..2803
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2822..3196
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3262..3509
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3278..3296)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3584..4172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3664..3683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, 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 ttcgaaacga ggaattcata atgtctgatg
      961 cggctccttc attgagcaat ctattttatg atccaacgta taatcctggt caaagcacca
     1021 ttaactacac ttccatatat gggaatggat ctaccatcac tttcgatgag ttgcaaggtt
     1081 tagttaacag tactgttact caggccatta tgtttggtgt cagatcaggc gccgctgctt
     1141 tgactttgat tgtcatgtgg atgacatcga gaagcagaaa aacgccgatt ttcattatca
     1201 accaagtttc attgttttta atcattttgc attctgcact ctattttaaa tatttactgt
     1261 ctaattactc ttcagtgact tacgctctca ccggatttcc tcagttcatc agtagaggtg
     1321 acgttcatgt ttatggtgct acaaatataa ttcaagtcct tcttgtggct tctattgaga
     1381 cttcactggt gtttcagata aaagttattt tcacaggcga caacttcaaa aggataggtt
     1441 tgatgctgac gtcgatatct ttcactttag ggattgctac agttaccatg tattttgtaa
     1501 gcgctgttaa aggtatgatt gtgacttata atgatgttag tgccacccaa gataaatact
     1561 tcaatgcatc cacaatttta cttgcatcct caataaactt tatgtcattt gtcctggtag
     1621 ttaaattgat tttagctatt agatcaagaa gattccttgg tctcaagcag ttcgatagtt
     1681 tccatatttt actcataatg tcagctcaat ctttgttggt accatcgata atattcatcc
     1741 tcgcatacag tttgaaacca aaccagggaa cagatgtctt gactactgtt gcaacattac
     1801 ttgctgtatt gtctttacca ttatcatcaa tgtgggccac ggctgctaat aatgcgtcga
     1861 ccaccgtctc caagacagag accagccaag tggcgcctgc ctaaggatcc taagcggccg
     1921 ccagcttggg cccgaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca
     1981 tcatcatcat catcattgag ttttagcctt agacatgact gttcctcagt tcaagttggg
     2041 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg
     2101 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
     2161 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
     2221 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
     2281 tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatcc
     2341 cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     2401 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     2461 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     2521 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     2581 tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
     2641 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
     2701 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagg gcggtgttga
     2761 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     2821 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     2881 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     2941 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     3001 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     3061 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     3121 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     3181 cgaggagcag gactgacacg tccgacgcgg cccgacgggt ccgaggcctc ggagatccgt
     3241 cccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc
     3301 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     3361 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     3421 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     3481 ttttgggacg ctcgaaggct ttaatttgca agctggagac caacatgtga gcaaaaggcc
     3541 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3601 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3661 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3721 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3781 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3841 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3901 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3961 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4021 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4081 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4141 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4201 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga tca
//
