LOCUS       sequence-224955-        5083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 5083)
  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: OsMATE
FEATURES             Location/Qualifiers
     source          1..5083
                     /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             2751..2774
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2775..2798
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2808..2825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2905..3151
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3166..3579
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3586..3633
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3652..4026
                     /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      4092..4339
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4108..4126)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4414..5002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4494..4513)
                     /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 ggaattcatg gcccgttctt
      961 ctagtgcacc tgagtctatg gaagctgacc atcaattaac tgcaactgtc actgccgccg
     1021 cctctggtga tatgcccgtt acagaacaac aacaacaaaa acagtgtatg gtcgcagttg
     1081 ccgccccccc agccactgag aatgccgatg cagctggcgg cggcggtgga gataacggtg
     1141 atcacttgcc acccacagct acctctttac ttggcggcgc aaggagaact ggcttacacc
     1201 ttttcgtgtt gaacgctcga agtgctctta ggttggatga attaggtgcc gaggtcctaa
     1261 gaattgcagt cccagcatct cttgctctga ctgcagatcc actagcatct ttaatagaca
     1321 cggcatttat tggacgtata ggtagtgttg agatagcagc tgtaggtgtt gcaattgctg
     1381 tttttaacca agtaatgaag gtctgcatat atccattagt ctcagttact acgtcattcg
     1441 tagccgaaga agacgctatc ttgtctaaag gtgctgctgg tgccgacgac gataacgatg
     1501 atggtcatga tgctaagggg catggagcat ctgctgctgc agtggcagac cccgagaaac
     1561 aacaagtggt cggtgtggac agtgcagaga ccaacggtgc tgaggtttca actgccgcag
     1621 tcagaacaac tgatgataag aaagccacag cagccggagt gggtgttggt aagtgcagaa
     1681 gaagatttgt accatccgtt acctctgctt taatagtcgg agcttttctt ggattattac
     1741 aggccgtttt tcttgttgct gctggtaaac ctttgttaag aattatgggc gttaagccag
     1801 gaagtccaat gatgattcca gctttgaggt atttagtcgt gagatcttta ggagcaccag
     1861 ctgtgttatt atcattggct atgcaaggag tcttcagagg ttttaaagac actaagacgc
     1921 cattgtatgc tactgtgacg ggtgatcttg ctaacatcgc tttagaccct attcttatat
     1981 tcacttgcag gtttggtgtc gttggtgcag ctatcgccca tgtcatttca cagtacctga
     2041 tcacacttat tatgctgtgt aagttggtca gaaaggttga tgttatacca agttcattga
     2101 aatcattaaa gttcagaaga tttttgggct gtggttttct attacttgcc agagttgtgg
     2161 ctgtgacttt ctgcgtgacc ttggccgctt cactggcagc gcgtcatggt gcaacagcta
     2221 tggctgcttt ccaaatttgt gcccaagtgt ggttagcctc atctttgttg gctgacggtc
     2281 tggctgttgc aggacaagct cttctggcct ctgctttcgc taagaaggac cactacaagg
     2341 tggctgttac aacagctaga gttttgcaac ttgctgttgt cctaggtgtt ggtttgacag
     2401 ccttcttagc tgctggtatg tggtttggag ctggcgtttt cacctcagat gctgccgtca
     2461 tctcaacaat acacagagga gtgccatttg tcgccggcac ccagactatt aatacattgg
     2521 ctttcgtttt tgacggtgtt aattttggtg catctgatta cgctttcgct gcctattcta
     2581 tggttggtgt tgctgccgtc actataccat gtttggtttt gcttagttct catggaggtt
     2641 tcgttggtat atggattgcc ttggcaattt atatgtctgt gagggctttc gctagtactt
     2701 ggagaatggg tgccgcccgt ggtccatgga aatttttgcg aaagggtacc ttggaagttt
     2761 tgttccaggg tccacatcat caccaccatc accatcattg agtcgaccat catcatcatc
     2821 atcattgagt ttgtagcctt agacatgact gttcctcagt tcaagttggg cacttacgag
     2881 aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat
     2941 gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt
     3001 cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata
     3061 tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac
     3121 tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc
     3181 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     3241 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     3301 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     3361 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     3421 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     3481 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     3541 cttgctcatt agaaagaaag catagcaatc taatctaagg gcggtgttga caattaatca
     3601 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     3661 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     3721 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     3781 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     3841 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     3901 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     3961 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     4021 gactgacacg tccgacgcgg cccgacgggt ccgaggcctc ggagatccgt cccccttttc
     4081 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     4141 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     4201 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     4261 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     4321 ctcgaaggct ttaatttgca agctggagac caacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4741 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga tca
//
