LOCUS       sequence-231199-        3794 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 3794)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3794)
  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: AtCOPT1 uPIC
FEATURES             Location/Qualifiers
     source          1..3794
                     /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             996..1025
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1493..1513
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1520..1537
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1617..1863
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1878..2291
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2298..2345
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2364..2738
                     /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      2804..3051
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2820..2838)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3126..3714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3206..3225)
                     /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 tatggatcat gatcacatgc atggaatgcc tcgaccatca tcatcatcat catcatcacc
     1021 atcatccatg atgaacaatg gatccatgaa cgaaggagga ggacatcacc acatgaagat
     1081 gatgatgcac atgaccttct tttggggcaa gaacacggag gttctcttct ccggttggcc
     1141 cggaacaagc tccggcatgt atgctctttg tcttatcttt gtcttctttc tcgccgtcct
     1201 caccgaatgg cttgctcatt cctctctcct ccgtggctcc accggagatt cggctaatcg
     1261 tgccgccggg ttaatccaaa ccgccgtgta taccctccgt atcggcctcg cctatctagt
     1321 catgctcgcc gtcatgtcgt ttaacgccgg tgtgtttctc gtcgctctgg ccggtcatgc
     1381 cgttggtttc atgttgttcg gaagccaaac tttccggaat acctccgatg accggaaaac
     1441 taactatgtt cctccctcag gttgtgcttg tgcattaaac ctcagcagcg gcgagaacct
     1501 gtacttccaa tctgtcgacc atcatcatca tcatcattga gtttgtagcc ttagacatga
     1561 ctgttcctca gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa
     1621 gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat
     1681 ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc
     1741 tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat
     1801 tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg
     1861 agaccttcgt ttgtgcggat cccccacaca ccatagcttc aaaatgtttc tactcctttt
     1921 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
     1981 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
     2041 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
     2101 aaatttttat cacgtttctt tttcttgaaa attttttttt ttgatttttt tctctttcga
     2161 tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca
     2221 tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa
     2281 tctaatctaa gggcggtgtt gacaattaat catcggcata gtatatcggc atagtataat
     2341 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     2401 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     2461 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     2521 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     2581 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     2641 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     2701 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtccgacgc ggcccgacgg
     2761 gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg atatcatgta attagttatg
     2821 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     2881 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     2941 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     3001 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctggag
     3061 accaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3781 ttggtcatga gatc
//
