LOCUS       sequence-231284-        4544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4544)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4544)
  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: OsPQL uPIC
FEATURES             Location/Qualifiers
     source          1..4544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             2243..2263
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2270..2287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2367..2613
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2628..3041
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3048..3095
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3114..3488
                     /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      3554..3801
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3570..3588)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3876..4464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3956..3975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcgc tgaggcttaa
      961 tatgggcatc ttcagtggag cgggagcaca tcgccctagc tgcccgtcgg ccgcgaattg
     1021 cgcaaaatgg gctcaaacct acctgaagta ctgcctttgt agcacgaggg atggcatggc
     1081 actgacgctt ggactgttaa gcgtcatcag ctggggagtt gctgaggtgc cacagatcat
     1141 aactaattac aagcataagt caacagaagg cctttccctt gcttttctaa tgacatggat
     1201 tgttggggac ttttttaacc ttattggctg cttcttggaa cctgagacgc tgccaacaca
     1261 attttacatg gctctgctat acaccatcac aactgttatt ctcactggac agacagtata
     1321 ctatagccac atatatcacc gtcttaaagc aaaaaaagcc agggcaacaa gcaagcctca
     1381 gaggcatcag cgtgctgatg cttcattgag agaaaagctt ttggggccca aggttattgg
     1441 ggaaatcaga aacaatagcc acctaggtgc aaccgtccct attccaacaa gttctccaat
     1501 tacagttaat acagaaatag tacggcaacg acatggccca agcagcctca gtgaatacta
     1561 ctacacgtca gcaagatctc tctcaagcag cccagtgcca atgtcaggca catggtcggc
     1621 taattatcat caaacaaatt ctccaccaga aatagatgac caaaaggaat ccttggtcag
     1681 tgagttttcc ccagcacaat atgcagcatc tcctttaata aagaattctc tctcagtggt
     1741 accatggatg tcccttctct tgggtatgag tgttttgcat tttttagttg gaacaaccca
     1801 tcaagaggtg ccaaatggta ttgtcatacc tgtcgggaga aggcttttac tgttggcgga
     1861 tgaccatgca gattcatctg tcagcaatgg cagtggaagt ggaattggaa gcttccttgg
     1921 ttgggctatg gcggtaatct atatgggtgg acgacttccc cagatctggt taaatatgaa
     1981 aagagggaat gctgagggac tcaatccact aatgtttact tttgcattag tgggaaatgt
     2041 gacatacgtt ggaagtatac ttgtgaaaag catggattgg tcaaaactta agccgaatct
     2101 accgtggctt gtagatgctg gaggatgtgt tttgcttgat acttttatta ttcttcagtt
     2161 tctttatttc cattatcgga agcgacacgt tcctgatgag cctgacagtg ctgataaagt
     2221 tgcattaaac ctcagcagcg gcgagaacct gtacttccaa tctgtcgacc atcatcatca
     2281 tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg
     2341 agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag
     2401 atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt
     2461 gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa
     2521 tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc
     2581 actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca
     2641 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     2701 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     2761 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     2821 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     2881 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
     2941 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
     3001 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat
     3061 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     3121 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     3181 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     3241 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     3301 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     3361 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     3421 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     3481 aggactgaca cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt
     3541 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     3601 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     3661 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3721 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     3781 cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag
     3841 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3901 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3961 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4021 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4081 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4141 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4201 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4261 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4321 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4381 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4441 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4501 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gatc
//
