LOCUS       sequence-230233-        4184 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 4184)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4184)
  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 3'
FEATURES             Location/Qualifiers
     source          1..4184
                     /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             1883..1903
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1910..1927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2007..2253
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2268..2681
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2688..2735
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2754..3128
                     /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      3194..3441
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3210..3228)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3516..4104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3596..3615)
                     /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 tatgctcctt gcctctaact ttgacactct ctcccacttc cttcaagatt ttggggacga
     1021 atattttgaa gcgagagaga agggggacta caaagtagtg gagaagagga aaaagaatct
     1081 tgagagatat aaaagtgttc ttgactccaa aagcgatgaa gaagctttgg ctaattacgc
     1141 agaatgggaa ccgcctcatg gacaatttag gtttaggcat ccatggaaac aatatgttgc
     1201 cgttggtgca ttactccggc agtgtgccta caggattgat gccttgaatt catacatcaa
     1261 ctcagatttt cagatcccag tggacataaa aaagaaatta gaaacaccat taagaagaat
     1321 gagctcggaa tcgggaaact caatgaaaga aatgtccatt tcactaaaac aaatgataaa
     1381 atcatcttct tctgatatcc atgtctcaaa ctcacaagct gcatgcaaat ctctttctac
     1441 tttacttaaa tcaggcatct tgaacgatgt tgagcctctt caaatgatct cattgatgac
     1501 gactgtctcc atgctcatag atattgttaa cttaaccgaa aagatctcgg aatctgtaca
     1561 cgagcttgca tccgctgcaa gatttaagaa caagatgagg ccgaccgtgc tatacgagaa
     1621 gtcggattcc ggaagcattg gtcgtgccat gccaatagat tctcatgaag atcatcatgt
     1681 tgtcacagtt ttacatgacg ttgataatga tagatcaaac aatgttgatg attcacgtgg
     1741 agggagttca caggactcat gtcatcatgt tgccataaag attgttgatg ataactcaaa
     1801 ccatgaaaaa catgaagatg gtgaaataca tgtacataca ttaagtaatg ggcatcttca
     1861 ggcattaaac ctcagcagcg gcgagaacct gtacttccaa tctgtcgacc atcatcatca
     1921 tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg
     1981 agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag
     2041 atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt
     2101 gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa
     2161 tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc
     2221 actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca
     2281 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     2341 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     2401 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     2461 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     2521 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
     2581 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
     2641 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat
     2701 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     2761 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     2821 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     2881 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     2941 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     3001 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     3061 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     3121 aggactgaca cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt
     3181 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     3241 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     3301 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3361 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     3421 cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag
     3481 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3541 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3601 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3661 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3721 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3781 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3841 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3901 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3961 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4021 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4081 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4141 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gatc
//
