LOCUS       sequence-267728-        4517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant form of 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 4517)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 4517)
  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: pPICZa-STE2-1D4
FEATURES             Location/Qualifiers
     source          1..4517
                     /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             join(940..996,997..1194,1195..1206)
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             2197..2226
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2242..2259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2339..2585
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2600..3013
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3020..3067
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3086..3460
                     /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      3526..3773
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3542..3560)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3928..3947)
                     /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 tgagatttcc ttcaattttt
      961 actgctgttt tattcgcagc atcctccgca ttagctgctc cagtcaacac tacaacagaa
     1021 gatgaaacgg cacaaattcc ggctgaagct gtcatcggtt actcagattt agaaggggat
     1081 ttcgatgttg ctgttttgcc attttccaac agcacaaata acgggttatt gtttataaat
     1141 actactattg ccagcattgc tgctaaagaa gaaggggtat ctctcgagaa aagagaggct
     1201 gaagctgaat tcataatgtc tgatgcggct ccttcattga gcaatctatt ttatgatcca
     1261 acgtataatc ctggtcaaag caccattaac tacacttcca tatatgggaa tggatctacc
     1321 atcactttcg atgagttgca aggtttagtt aacagtactg ttactcaggc cattatgttt
     1381 ggtgtcagat caggcgccgc tgctttgact ttgattgtca tgtggatgac atcgagaagc
     1441 agaaaaacgc cgattttcat tatcaaccaa gtttcattgt ttttaatcat tttgcattct
     1501 gcactctatt ttaaatattt actgtctaat tactcttcag tgacttacgc tctcaccgga
     1561 tttcctcagt tcatcagtag aggtgacgtt catgtttatg gtgctacaaa tataattcaa
     1621 gtccttcttg tggcttctat tgagacttca ctggtgtttc agataaaagt tattttcaca
     1681 ggcgacaact tcaaaaggat aggtttgatg ctgacgtcga tatctttcac tttagggatt
     1741 gctacagtta ccatgtattt tgtaagcgct gttaaaggta tgattgtgac ttataatgat
     1801 gttagtgcca cccaagataa atacttcaat gcatccacaa ttttacttgc atcctcaata
     1861 aactttatgt catttgtcct ggtagttaaa ttgattttag ctattagatc aagaagattc
     1921 cttggtctca agcagttcga tagtttccat attttactca taatgtcagc tcaatctttg
     1981 ttggtaccat cgataatatt catcctcgca tacagtttga aaccaaacca gggaacagat
     2041 gtcttgacta ctgttgcaac attacttgct gtattgtctt taccattatc atcaatgtgg
     2101 gccacggctg ctaataatgc gtcgaccacc gtctccaaga cagagaccag ccaagtggcg
     2161 cctgcctaag gatcctaagc ggccgccagc tttctagaac aaaaactcat ctcagaagag
     2221 gatctgaata gcgccgtcga ccatcatcat catcatcatt gagtttgtag ccttagacat
     2281 gactgttcct cagttcaagt tgggcactta cgagaagacc ggtcttgcta gattctaatc
     2341 aagaggatgt cagaatgcca tttgcctgag agatgcaggc ttcatttttg atactttttt
     2401 atttgtaacc tatatagtat aggatttttt ttgtcatttt gtttcttctc gtacgagctt
     2461 gctcctgatc agcctatctc gcagctgatg aatatcttgt ggtaggggtt tgggaaaatc
     2521 attcgagttt gatgtttttc ttggtatttc ccactcctct tcagagtaca gaagattaag
     2581 tgagaccttc gtttgtgcgg atcccccaca caccatagct tcaaaatgtt tctactcctt
     2641 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     2701 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     2761 aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     2821 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc
     2881 gatgacctcc cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt
     2941 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc
     3001 aatctaatct aagggcggtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3061 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     3121 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3181 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3241 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3301 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3361 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3421 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtccgac gcggcccgac
     3481 gggtccgagg cctcggagat ccgtccccct tttcctttgt cgatatcatg taattagtta
     3541 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     3601 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     3661 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     3721 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcaagctgg
     3781 agaccaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagatca
//
