LOCUS       sequence-231192-        3719 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 3719)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3719)
  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: AtCOPT6 uPIC
FEATURES             Location/Qualifiers
     source          1..3719
                     /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             981..998
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1418..1438
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1445..1462
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1542..1788
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1803..2216
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2223..2270
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2289..2663
                     /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      2729..2976
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2745..2763)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3131..3150)
                     /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 ggtaacatgc caccatcatc accatcatca atggtaaatc ataccaattc
     1021 caacatgatc atgatgcata tgactttctt ctggggaaag aacacggaga tactcttctc
     1081 cggctggccc gggacaagtc tcggcatgta cgttctatgt ctcatcgtcg tcttcctcct
     1141 tgctgttatc gtcgagtggc ttgcccattc ttcgatcctt cgtggacgtg gctccactag
     1201 ccgcgccaag gggctcgttc agaccgcagt ctacacactc aagacaggcc ttgcttatct
     1261 cgtgatgctc gccgttatgt cctttaacgg tggagttttc attgtcgcta tcgccggatt
     1321 cgccgtgggt ttcatgctct tcggaagcac tgctttcaag aacccctctg atgacgagaa
     1381 acccttcgaa caactagcat taaacctcag cagcggcgag aacctgtact tccaatctgt
     1441 cgaccatcat catcatcatc attgagtttg tagccttaga catgactgtt cctcagttca
     1501 agttgggcac ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg
     1561 ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag
     1621 tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat
     1681 ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt
     1741 ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg
     1801 cggatccccc acacaccata gcttcaaaat gtttctactc cttttttact cttccagatt
     1861 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     1921 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     1981 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     2041 ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc tcccattgat
     2101 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
     2161 tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa tctaagggcg
     2221 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     2281 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     2341 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     2401 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     2461 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     2521 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     2581 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     2641 tcgtggccga ggagcaggac tgacacgtcc gacgcggccc gacgggtccg aggcctcgga
     2701 gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     2761 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     2821 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     2881 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     2941 gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca
     3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatc
//
