LOCUS       sequence-230350-        3866 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 3866)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3866)
  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: AtALMT1-uPIC 5'
FEATURES             Location/Qualifiers
     source          1..3866
                     /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             complement(1067..1078)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1565..1585
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1592..1609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1689..1935
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1950..2363
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2370..2417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2436..2810
                     /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      2876..3123
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2892..2910)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3198..3786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3278..3297)
                     /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 tatggagaaa gtgagagaga tagtgagaga agggattaga gtagggaatg aagacccgag
     1021 aagaattatt catgctttca aagtgggact tgctcttgtt ttggtctctt ccttctatta
     1081 ttaccaacct ttcggtcctt tcaccgatta cttcggtatc aatgcaatgt gggctgtcat
     1141 gaccgtcgtt gttgtctttg aattctccgt aggggctaca cttgggaaag gactaaatag
     1201 aggagtggca acattagtag caggaggact aggaattgga gctcatcaac tagcaagatt
     1261 gtcgggtgca acagtggaac ctattctttt ggtcatgttg gtctttgtgc aagctgcgtt
     1321 gtcgacgttc gtaaggtttt tcccatgggt aaagacaaag ttcgattatg ggatattgat
     1381 attcatattg acgttcgcgc tgatatcgtt gtcggggttt agagacgaag agataatgga
     1441 tttggcagaa agcagactat cgacagtggt gattggagga gtcagttgca tccttatctc
     1501 tatctttgtc tgtcctgttt gggctggaca agatctacat tctgcattaa acctcagcag
     1561 cggcgagaac ctgtacttcc aatctgtcga ccatcatcat catcatcatt gagtttgtag
     1621 ccttagacat gactgttcct cagttcaagt tgggcactta cgagaagacc ggtcttgcta
     1681 gattctaatc aagaggatgt cagaatgcca tttgcctgag agatgcaggc ttcatttttg
     1741 atactttttt atttgtaacc tatatagtat aggatttttt ttgtcatttt gtttcttctc
     1801 gtacgagctt gctcctgatc agcctatctc gcagctgatg aatatcttgt ggtaggggtt
     1861 tgggaaaatc attcgagttt gatgtttttc ttggtatttc ccactcctct tcagagtaca
     1921 gaagattaag tgagaccttc gtttgtgcgg atcccccaca caccatagct tcaaaatgtt
     1981 tctactcctt ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca
     2041 aaacacccaa gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt
     2101 acccgtacta aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga
     2161 aaaaggcaat aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt
     2221 tttctctttc gatgacctcc cattgatatt taagttaata aacggtcttc aatttctcaa
     2281 gtttcagttt catttttctt gttctattac aacttttttt acttcttgct cattagaaag
     2341 aaagcatagc aatctaatct aagggcggtg ttgacaatta atcatcggca tagtatatcg
     2401 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     2461 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     2521 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     2581 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     2641 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     2701 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     2761 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtccgac
     2821 gcggcccgac gggtccgagg cctcggagat ccgtccccct tttcctttgt cgatatcatg
     2881 taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag
     2941 gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta
     3001 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca
     3061 tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaatt
     3121 tgcaagctgg agaccaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3601 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3841 cgttaaggga ttttggtcat gagatc
//
