LOCUS       sequence-267651-        3809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast STE2 coding sequence in vector for protein expression in the 
            methanogenic yeast Pichia pastoris, with GAP promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3809)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 3809)
  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: pGAPZ-STE2-1D4
FEATURES             Location/Qualifiers
     source          1..3809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..477
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             1481..1510
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1526..1543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1622..1868
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1883..2296
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2303..2350
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2369..2743
                     /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      2809..3056
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2825..2843)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3131..3719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3211..3230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttgtaga aatgtcttgg tgtcctcgtc caatcaggta gccatctctg aaatatctgg
       61 ctccgttgca actccgaacg acctgctggc aacgtaaaat tctccggggt aaaacttaaa
      121 tgtggagtaa tggaaccaga aacgtctctt cccttctctc tccttccacc gcccgttacc
      181 gtccctagga aattttactc tgctggagag cttcttctac ggcccccttg cagcaatgct
      241 cttcccagca ttacgttgcg ggtaaaacgg aggtcgtgta cccgacctag cagcccaggg
      301 atggaaaagt cccggccgtc gctggcaata atagcgggcg gacgcatgtc atgagattat
      361 tggaaaccac cagaatcgaa tataaaaggc gaacaccttt cccaattttg gtttctcctg
      421 acccaaagac tttaaattta atttatttgt ccctatttca atcaattgaa caactatttc
      481 gaaacgagga attcataatg tctgatgcgg ctccttcatt gagcaatcta ttttatgatc
      541 caacgtataa tcctggtcaa agcaccatta actacacttc catatatggg aatggatcta
      601 ccatcacttt cgatgagttg caaggtttag ttaacagtac tgttactcag gccattatgt
      661 ttggtgtcag atcaggcgcc gctgctttga ctttgattgt catgtggatg acatcgagaa
      721 gcagaaaaac gccgattttc attatcaacc aagtttcatt gtttttaatc attttgcatt
      781 ctgcactcta ttttaaatat ttactgtcta attactcttc agtgacttac gctctcaccg
      841 gatttcctca gttcatcagt agaggtgacg ttcatgttta tggtgctaca aatataattc
      901 aagtccttct tgtggcttct attgagactt cactggtgtt tcagataaaa gttattttca
      961 caggcgacaa cttcaaaagg ataggtttga tgctgacgtc gatatctttc actttaggga
     1021 ttgctacagt taccatgtat tttgtaagcg ctgttaaagg tatgattgtg acttataatg
     1081 atgttagtgc cacccaagat aaatacttca atgcatccac aattttactt gcatcctcaa
     1141 taaactttat gtcatttgtc ctggtagtta aattgatttt agctattaga tcaagaagat
     1201 tccttggtct caagcagttc gatagtttcc atattttact cataatgtca gctcaatctt
     1261 tgttggtacc atcgataata ttcatcctcg catacagttt gaaaccaaac cagggaacag
     1321 atgtcttgac tactgttgca acattacttg ctgtattgtc tttaccatta tcatcaatgt
     1381 gggccacggc tgctaataat gcgtcgacca ccgtctccaa gacagagacc agccaagtgg
     1441 cgcctgccta aggatcctaa gcggccgcca gcttgggccc gaacaaaaac tcatctcaga
     1501 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt tagccttaga
     1561 catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg ctagattcta
     1621 atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt
     1681 tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag
     1741 cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa
     1801 atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt
     1861 aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat gtttctactc
     1921 cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc
     1981 caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta
     2041 ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc
     2101 aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct
     2161 ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag
     2221 tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat
     2281 agcaatctaa tctaagggcg gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     2341 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
     2401 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     2461 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     2521 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     2581 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     2641 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     2701 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc gacgcggccc
     2761 gacgggtccg aggcctcgga gatccgtccc ccttttcctt tgtcgatatc atgtaattag
     2821 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     2881 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     2941 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     3001 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
     3061 tggagaccaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3121 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3181 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3241 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3301 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3361 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3421 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3481 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3541 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3601 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3661 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3781 ggattttggt catgcatgag atcagatct
//
