LOCUS       sequence-230228-        3989 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 3989)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3989)
  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 3'
FEATURES             Location/Qualifiers
     source          1..3989
                     /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             1688..1708
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1715..1732
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1812..2058
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2073..2486
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2493..2540
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2559..2933
                     /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      2999..3246
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3015..3033)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3321..3909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3401..3420)
                     /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 tatgctcgcc tccggcaacc tcgacaaact cgctcagttc attgaaggaa tggaattcaa
     1021 ctgctttggc gaaaacagtg ttgcaaataa ttttggggga aaagatttcc cccaaatgca
     1081 caagagcgtc cttaattcga aggccactga ggactctttg tgcacctttg ccaaatggga
     1141 gcctcgtcat ggccagttca gatttcgaca cccatggagt caataccaga agctgggaac
     1201 tctttgtcgc caatgtgcgt cttctatgga ggctcttgct tcatatgtca tcacaacctc
     1261 aaaaacccag tgccctgctg cagccaaccc tgagctatca tgtaaggttc gaaaaacatg
     1321 tggcgaaatg agcttgcatt cctccaaggt gcttagggat ctcgcaatgg caactcgaac
     1381 aatgactgtg ccgtctccag tgaatatcac catggctaca gccgtgaaag cagcggaaag
     1441 cctcagaagc gagcttgcag agaacacggc tctgttgcaa gtgatgcatg tggccgtcac
     1501 cgcaacactt cttgcggact tggttgatag ggtgaaggaa atcgcggaat gtgttgatgt
     1561 cctagcaaga ctggcgcact ttaagaaccc cgaggacaca aaaaatgtcg ttgttagtac
     1621 cgtgagtcga gggatagacg aacctttgcc tgacgtggtt attttgccat taaacctcag
     1681 cagcggcgag aacctgtact tccaatctgt cgaccatcat catcatcatc attgagtttg
     1741 tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg
     1801 ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt
     1861 ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt
     1921 ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg
     1981 gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt
     2041 acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat
     2101 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     2161 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     2221 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     2281 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     2341 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     2401 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     2461 aagaaagcat agcaatctaa tctaagggcg gtgttgacaa ttaatcatcg gcatagtata
     2521 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     2581 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     2641 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     2701 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     2761 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     2821 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     2881 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc
     2941 gacgcggccc gacgggtccg aggcctcgga gatccgtccc ccttttcctt tgtcgatatc
     3001 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
     3061 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     3121 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     3181 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta
     3241 atttgcaagc tggagaccaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3961 tcacgttaag ggattttggt catgagatc
//
