LOCUS       sequence-231165-        4661 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 4661)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4661)
  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: TaALMT-uPIC
FEATURES             Location/Qualifiers
     source          1..4661
                     /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             2360..2380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2387..2404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2484..2730
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2745..3158
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3165..3212
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3231..3605
                     /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      3671..3918
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3687..3705)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /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 cacaagctcg cctccggcaa cctcgacaaa ctcgctcagt tcattgaagg
     1681 aatggaattc aactgctttg gcgaaaacag tgttgcaaat aattttgggg gaaaagattt
     1741 cccccaaatg cacaagagcg tccttaattc gaaggccact gaggactctt tgtgcacctt
     1801 tgccaaatgg gagcctcgtc atggccagtt cagatttcga cacccatgga gtcaatacca
     1861 gaagctggga actctttgtc gccaatgtgc gtcttctatg gaggctcttg cttcatatgt
     1921 catcacaacc tcaaaaaccc agtgccctgc tgcagccaac cctgagctat catgtaaggt
     1981 tcgaaaaaca tgtggcgaaa tgagcttgca ttcctccaag gtgcttaggg atctcgcaat
     2041 ggcaactcga acaatgactg tgccgtctcc agtgaatatc accatggcta cagccgtgaa
     2101 agcagcggaa agcctcagaa gcgagcttgc agagaacacg gctctgttgc aagtgatgca
     2161 tgtggccgtc accgcaacac ttcttgcgga cttggttgat agggtgaagg aaatcgcgga
     2221 atgtgttgat gtcctagcaa gactggcgca ctttaagaac cccgaggaca caaaaaatgt
     2281 cgttgttagt accgtgagtc gagggataga cgaacctttg cctgacgtgg ttattttgcc
     2341 attaaacctc agcagcggcg agaacctgta cttccaatct gtcgaccatc atcatcatca
     2401 tcattgagtt tgtagcctta gacatgactg ttcctcagtt caagttgggc acttacgaga
     2461 agaccggtct tgctagattc taatcaagag gatgtcagaa tgccatttgc ctgagagatg
     2521 caggcttcat ttttgatact tttttatttg taacctatat agtataggat tttttttgtc
     2581 attttgtttc ttctcgtacg agcttgctcc tgatcagcct atctcgcagc tgatgaatat
     2641 cttgtggtag gggtttggga aaatcattcg agtttgatgt ttttcttggt atttcccact
     2701 cctcttcaga gtacagaaga ttaagtgaga ccttcgtttg tgcggatccc ccacacacca
     2761 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     2821 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     2881 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     2941 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     3001 tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt taataaacgg
     3061 tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc
     3121 ttgctcatta gaaagaaagc atagcaatct aatctaaggg cggtgttgac aattaatcat
     3181 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
     3241 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
     3301 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
     3361 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
     3421 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
     3481 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
     3541 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
     3601 actgacacgt ccgacgcggc ccgacgggtc cgaggcctcg gagatccgtc ccccttttcc
     3661 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     3721 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     3781 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     3841 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     3901 tcgaaggctt taatttgcaa gctggagacc aacatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgaaa actcacgtta agggattttg gtcatgagat c
//
