LOCUS       sequence-230349-        3959 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 3959)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3959)
  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: TaALMT1-uPIC 5'
FEATURES             Location/Qualifiers
     source          1..3959
                     /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             1658..1678
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1685..1702
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1782..2028
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2043..2456
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2463..2510
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2529..2903
                     /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      2969..3216
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2985..3003)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3291..3879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3371..3390)
                     /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 tatggatatt gatcacggca gagagagcga cggcgagatg gtgggcacca tcgccagctg
     1021 cgggctgctg ctccactcgc ttctcgccgg gctcgggcgt cgcgccgccg ggttcgcccg
     1081 gaaggtgggc ggcgccgcgc gggaggaccc gaggcgggtg gcgcactcgc tcaaagtcgg
     1141 cctggcgctc gcgctggtgt ccgtcgtcta cttcgtcacg ccgctcttca acggcctcgg
     1201 ggtgtccgcg atatgggccg tgctcaccgt cgtcgtcgtc atggagtaca ccgtcggtgc
     1261 cacgctgagt aaaggcttga acagagcctt ggcgacgttg gtggctggct gcatcgccgt
     1321 cggagctcat cagttagctg aattagctga acgctgtggt gatcagggag agcccataat
     1381 gcttaccgtg ctcgtcttct tcgtagcgtc agcggcgacg ttcttgcgct tcatcccgga
     1441 gatcaaggcc aagtacgact acggcgtgac catcttcata ctgaccttcg gtctggtggc
     1501 cgtgtcgagc tacagagtgg aggagctcat ccagctcgcg caccagcggt tctacaccat
     1561 agccgtcggc gtcttcatct gcctctgcac caccgtcttc ctcttccccg tctgggccgg
     1621 agaggacgtc cacaaggcat taaacctcag cagcggcgag aacctgtact tccaatctgt
     1681 cgaccatcat catcatcatc attgagtttg tagccttaga catgactgtt cctcagttca
     1741 agttgggcac ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg
     1801 ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag
     1861 tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat
     1921 ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt
     1981 ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg
     2041 cggatccccc acacaccata gcttcaaaat gtttctactc cttttttact cttccagatt
     2101 ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc atactaaatt
     2161 tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa
     2221 aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt
     2281 ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc tcccattgat
     2341 atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt cttgttctat
     2401 tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa tctaagggcg
     2461 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     2521 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     2581 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     2641 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     2701 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     2761 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     2821 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     2881 tcgtggccga ggagcaggac tgacacgtcc gacgcggccc gacgggtccg aggcctcgga
     2941 gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     3001 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     3061 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     3121 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     3181 gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca
     3241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatc
//
