LOCUS       sequence-256401-        3792 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 3792)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 3792)
  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..3792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(96..115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        695..1171
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2160..2189
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2205..2222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2301..2547
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2562..2973
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2980..3027
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3046..3420
                     /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      3486..3733
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3502..3520)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
       61 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      181 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
      241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      421 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      661 ttaagggatt ttggtcatgc atgagatcag atcttttttg tagaaatgtc ttggtgtcct
      721 cgtccaatca ggtagccatc tctgaaatat ctggctccgt tgcaactccg aacgacctgc
      781 tggcaacgta aaattctccg gggtaaaact taaatgtgga gtaatggaac cagaaacgtc
      841 tcttcccttc tctctccttc caccgcccgt taccgtccct aggaaatttt actctgctgg
      901 agagcttctt ctacggcccc cttgcagcaa tgctcttccc agcattacgt tgcgggtaaa
      961 acggaggtcg tgtacccgac ctagcagccc agggatggaa aagtcccggc cgtcgctggc
     1021 aataatagcg ggcggacgca tgtcatgaga ttattggaaa ccaccagaat cgaatataaa
     1081 aggcgaacac ctttcccaat tttggtttct cctgacccaa agactttaaa tttaatttat
     1141 ttgtccctat ttcaatcaat tgaacaacta tttcgaaacg aggaattcat gtctgatgcg
     1201 gctccttcat tgagcaatct attttatgat ccaacgtata atcctggtca aagcaccatt
     1261 aactacactt ccatatatgg gaatggatct accatcactt tcgatgagtt gcaaggttta
     1321 gttaacagta ctgttactca ggccattatg tttggtgtca gatgtggtgc agctgctttg
     1381 actttgattg tcatgtggat gacatcgaga agcagaaaaa cgccgatttt cattatcaac
     1441 caagtttcat tgtttttaat cattttgcat tctgcactct attttaaata tttactgtct
     1501 aattactctt cagtgactta cgctctcacc ggatttcctc agttcatcag tagaggtgac
     1561 gttcatgttt atggtgctac aaatataatt caagtccttc ttgtggcttc tattgagact
     1621 tcactggtgt ttcagataaa agttattttc acaggcgaca acttcaaaag gataggtttg
     1681 atgctgacgt cgatatcttt cactttaggg attgctacag ttaccatgta ttttgtaagc
     1741 gctgttaaag gtatgattgt gacttataat gatgttagtg ccacccaaga taaatacttc
     1801 aatgcatcca caattttact tgcatcctca ataaacttta tgtcatttgt cctggtagtt
     1861 aaattgattt tagctattag atcaagaaga ttccttggtc tcaagcagtt cgatagtttc
     1921 catattttac tcataatgtc atgtcaatct ttgttggttc catcgataat attcatcctc
     1981 gcatacagtt tgaaaccaaa ccagggaaca gatgtcttga ctactgttgc aacattactt
     2041 gctgtattgt ctttaccatt atcatcaatg tgggccacgg ctgctaataa tgcgtcgacc
     2101 accgtctcca agacagagac cagccaagtg gcgcctgccg cggccgccag cttnnnnnng
     2161 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
     2221 attgagtttt agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
     2281 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     2341 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     2401 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     2461 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     2521 cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatccccca cacaccatag
     2581 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
     2641 cgtaccactt caaaacaccc aagcacagca tactaaattt tccctctttc ttcctctagg
     2701 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
     2761 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaatttttt
     2821 tttttagttt ttttctcttt cagtgacctc cattgatatt taagttaata aacggtcttc
     2881 aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt acttcttgtt
     2941 cattagaaag aaagcatagc aatctaatct aagggcggtg ttgacaatta atcatcggca
     3001 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     3061 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     3121 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     3181 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     3241 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     3301 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     3361 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     3421 cacgtccgac ggcggcccac gggtcccagg cctcggagat ccgtccccct tttcctttgt
     3481 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     3541 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     3601 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     3661 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     3721 ggctttaatt tgcaagctgg agaccaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3781 ccgtaaaaag gc
//
