LOCUS       sequence-230230-        4148 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 4148)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4148)
  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: AtPQL uPIC
FEATURES             Location/Qualifiers
     source          1..4148
                     /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             1847..1867
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1874..1891
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1971..2217
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2232..2645
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2652..2699
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2718..3092
                     /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      3158..3405
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3174..3192)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /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 tatgattcga gatgatttgt cactatctct tggtatcatc agtgttatca gttggagtgt
     1021 tgccgagata ccgcagatta tgactaacta caatcaaaag tccattgaag gagtctctat
     1081 aaccttctta acaacatgga tgcttggtga tatctttaat gttgttggct gcttgatgga
     1141 accagctagt cttccagttc aattctacac ggcagtgttg tatactctgg ctacacttgt
     1201 tctctatgtt caatctatat actacgggca tatttacccg cggttgatga aaaaccgaag
     1261 gaatcatcag atggttgatg tagaagagcc tttgcttcgt gaagaagcta agcgtccttc
     1321 taccaaatcg ttgctatgtg tggtctcggt gttcttgttt cttggatcat tcaacgtatt
     1381 gagtggttca agaagtatgg atttgagagg gaaagataga gtgtttgtag ttggaggagc
     1441 aggagcaaga aagcttttgg aggtctctag cggtaactta ggagaaaaca acaatattgg
     1501 aatgtggttg ggttgggcaa tggcggcgat ttacatgggt ggaaggcttc ctcaaatatg
     1561 tatgaatgta aggagaggaa atgttgaggg attgaatcca ttaatgttct tctttgcatt
     1621 cattggcaat gtgacttatg ttgcaagcat actagtgaac agcgtcgaat ggtcgaaaat
     1681 cgaaccgaat ctaccatggc tagttgattc aggaggatgt gctgttcttg attttcttat
     1741 tctgcttcag tttttctact ttcactgtcg caaagtcgag gcagattccg tcaagaagaa
     1801 gcatgaaacc ggtgaagaag ctgtcgcatt aaacctcagc agcggcgaga acctgtactt
     1861 ccaatctgtc gaccatcatc atcatcatca ttgagtttgt agccttagac atgactgttc
     1921 ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat
     1981 gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa
     2041 cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga
     2101 tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt
     2161 ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct
     2221 tcgtttgtgc ggatccccca cacaccatag cttcaaaatg tttctactcc ttttttactc
     2281 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca
     2341 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg
     2401 gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt
     2461 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct
     2521 cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc
     2581 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat
     2641 ctaagggcgg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2701 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     2761 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     2821 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     2881 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     2941 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     3001 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     3061 gcgtgcactt cgtggccgag gagcaggact gacacgtccg acgcggcccg acgggtccga
     3121 ggcctcggag atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
     3181 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     3241 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     3301 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     3361 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagatc
//
