LOCUS       sequence-231287-        3758 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 3758)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3758)
  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: AtCOPT2 uPIC
FEATURES             Location/Qualifiers
     source          1..3758
                     /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             990..1013
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1457..1477
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1484..1501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1581..1827
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1842..2255
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2262..2309
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2328..2702
                     /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      2768..3015
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2784..2802)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3090..3678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3170..3189)
                     /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 atgacatgcc accaccatca ccatcatcat catcgatgtc
     1021 gaatcataca acaccacaca tgatgatgat gcacatgacc ttcttttggg gtaagaacac
     1081 ggaggttctc ttctccggat ggcccgggac aagctccggc atgtacgctc tctgtctcat
     1141 tgttatattc ctcctcgccg taattgccga gtggctcgct cattcaccga tcctacgtgt
     1201 cagtggctca accaatcgag ccgccgggct cgctcaaacc gctgtgtaca cactcaagac
     1261 tggcctttcg tatttggtga tgctcgctgt tatgtccttt aacgcaggtg ttttcatcgt
     1321 cgctatcgcc ggctatggcg ttggtttctt tctctttggg agcactactt tcaagaaacc
     1381 ctccgatgac cagaaaaccg ccgagcttct tccgccgtct tcaggctgcg tttgtgcatt
     1441 aaacctcagc agcggcgaga acctgtactt ccaatctgtc gaccatcatc atcatcatca
     1501 ttgagtttgt agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga
     1561 ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag
     1621 gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt
     1681 ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt
     1741 gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct
     1801 cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatccccca cacaccatag
     1861 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
     1921 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg
     1981 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
     2041 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt
     2101 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct
     2161 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg
     2221 ctcattagaa agaaagcata gcaatctaat ctaagggcgg tgttgacaat taatcatcgg
     2281 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     2341 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     2401 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     2461 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     2521 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     2581 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     2641 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     2701 gacacgtccg acgcggcccg acgggtccga ggcctcggag atccgtcccc cttttccttt
     2761 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
     2821 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     2881 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     2941 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     3001 aaggctttaa tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg
     3061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3721 aacgaaaact cacgttaagg gattttggtc atgagatc
//
