LOCUS       sequence-220657-        4881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4881)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4881)
  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: MtPT4
FEATURES             Location/Qualifiers
     source          1..4881
                     /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             2563..2592
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2608..2625
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2705..2951
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2966..3377
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3385..3432
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3451..3825
                     /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      3891..4138
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3907..3925)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4213..4801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4293..4312)
                     /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 aggaattcat gggattagaa
      961 gtccttgagg ctcttgattc agctcgtaca caatggtacc atgtaacagc tatagtaatt
     1021 gctggcatgg gattcttcac tgatgcctat gatctgttct gcatttcaac cgtttcgaag
     1081 ctcttgggac gattatatta ctttgatcca agcactaaca aaccagggaa gcttccacca
     1141 tcagttaata atgtagtcac cggtgtggcc cttgtcggaa cactttctgg ccaattagtc
     1201 ttcggctggc tcggagacaa acttggacgc aagaaagttt atggtgtcac actcattatc
     1261 atggttgctt gtgccatttg ctctggtctt tcttttggtt cttccgcgaa atctgttatg
     1321 ataacccttt gttttttcag gttttggctt ggatttggta ttggtggaga ctacccttta
     1381 tcagcaacca tcatgtctga atacgccaac aaaaggacac gaggcgcttt catagcagca
     1441 gttttcgcaa tgcaaggcgt gggaatcatc tttgccggat tggtttcaat ggtattttca
     1501 ggaattttta aagcatacta ccaggctccc cgtttcaatg aagacccaat tttatcaaca
     1561 cagcccgaag gggatttact ctggcgatta attctcatga tcggtgctgt tccagctgcg
     1621 atgacttact actggagaat gaaaatgcct gaaaccggtc gttacactgc aatcgtagaa
     1681 gggaacgcga aacaagccgc tgcagacatg gcaagagttt tggacatcga aatcatagcg
     1741 gagcaagata agttggctga atttaaagca gcaaatgatt accctctttg gtctagcgag
     1801 tttttcaaca ggcacggtcg tcatttaatc ggtacaatga gttgttggtt cttgttagat
     1861 attgctttct acagtcagaa tttgacacag aaagatattt accctgcaat gggacttatt
     1921 cgtcaagaca aagaaatgaa tgctattgat gaagttttcc aaacttcgcg cgccatgttt
     1981 gttgtcgcct tgtttggaac attccccggt tattggttca ctgttttctt cattgaaaag
     2041 cttggaagat tcaagataca gttggttggt ttcttcatga tgtctttctt catgtttgtc
     2101 attggtgtta aatatgaata cttgaaagac gaaaacaaaa acctcttcgc tcttttatac
     2161 gggctaacat tcttcttcgc gaactttggg cctaatagta caacttttgt gttgcccgct
     2221 gagctttttc ctacgcgggt gaggtcaaca tgccatgctt tcagtgctgc atctgggaag
     2281 gcgggtgcta tggttggtgc gttcgggata caatactaca cgttggatgg tacgccaagg
     2341 aagattagac gcgcaatgat gattctggct tttacaaatt tgataggatt cttttgcacg
     2401 ttcttggtga cggaaactaa gggacgatct ttggaagaga tttcgggtga ggatggaaga
     2461 gagagtgaac tcactgcaac accaaatgac agggctccgg ggattcgcca ggactcaaga
     2521 actgagaaga tgctcgagcc gcggcggccg ccagcttacg tagaacaaaa actcatctca
     2581 gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttgtagcctt
     2641 agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc ttgctagatt
     2701 ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac
     2761 ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt cttctcgtac
     2821 gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta ggggtttggg
     2881 aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag agtacagaag
     2941 attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa aatgtttcta
     3001 ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac
     3061 acccaagcac agcatactaa attttccctc tttcttcctc tagggtgtcg ttaattaccc
     3121 gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa
     3181 ggcaataaaa atttttatca cgtttctttt tcttgaaatt ttttttttta gtttttttct
     3241 ctttcagtga cctccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     3301 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgttcattag aaagaaagca
     3361 tagcaatcta atctaagggg cggtgttgac aattaatcat cggcatagta tatcggcata
     3421 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     3481 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     3541 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     3601 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3661 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3721 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3781 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt ccgacggcgg
     3841 cccacgggtc ccaggcctcg gagatccgtc ccccttttcc tttgtcgata tcatgtaatt
     3901 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     3961 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     4021 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     4081 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa
     4141 gctggagacc aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4381 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     4441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4621 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4861 agggattttg gtcatgagat c
//
