LOCUS       sequence-225001-        4882 bp    DNA     circular SYN 14-APR-2025
DEFINITION  protein expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4882)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..4882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..939
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             2562..2591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2607..2624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2704..2950
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2965..3378
                     /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 gatctaacat ccaaagacga aaggttgaat gaaacctttt tgccatccga catccacagg
       61 tccattctca cacataagtg ccaaacgcaa caggagggga tacactagca gcagaccgtt
      121 gcaaacgcag gacctccact cctcttctcc tcaacaccca cttttgccat cgaaaaacca
      181 gcccagttat tgggcttgat tggagctcgc tcattccaat tccttctatt aggctactaa
      241 caccatgact ttattagcct gtctatcctg gcccccctgg cgaggttcat gtttgtttat
      301 ttccgaatgc aacaagctcc gcattacacc cgaacatcac tccagatgag ggctttctga
      361 gtgtggggtc aaatagtttc atgttcccca aatggcccaa aactgacagt ttaaacgctg
      421 tcttggaacc taatatgaca aaagcgtgat ctcatccaag atgaactaag tttggttcgt
      481 tgaaatgcta acggccagtt ggtcaaaaag aaacttccaa aagtcggcat accgtttgtc
      541 ttgtttggta ttgattgacg aatgctcaaa aataatctca ttaatgctta gcgcagtctc
      601 tctatcgctt ctgaaccccg gtgcacctgt gccgaaacgc aaatggggaa acacccgctt
      661 tttggatgat tatgcattgt ctccacattg tatgcttcca agattctggt gggaatactg
      721 ctgatagcct aacgttcatg atcaaaattt aactgttcta acccctactt gacagcaata
      781 tataaacaga aggaagctgc cctgtcttaa accttttttt ttatcatcat tattagctta
      841 ctttcataat tgcgactggt tccaattgac aagcttttga ttttaacgac ttttaacgac
      901 aacttgagaa gatcaaaaaa caactaatta ttcgaaacga ggaattcatg ggattagaag
      961 tccttgaggc tcttgattca gctcgtacac aatggtacca tgtaacagct atagtaattg
     1021 ctggcatggg attcttcact gatgcctatg atctgttctg catttcaacc gtttcgaagc
     1081 tcttgggacg attatattac tttgatccaa gcactaacaa accagggaag cttccaccat
     1141 cagttaataa tgtagtcacc ggtgtggccc ttgtcggaac actttctggc caattagtct
     1201 tcggctggct cggagacaaa cttggacgca agaaagttta tggtgtcaca ctcattatca
     1261 tggttgcttg tgccatttgc tctggtcttt cttttggttc ttccgcgaaa tctgttatga
     1321 taaccctttg ttttttcagg ttttggcttg gatttggtat tggtggagac taccctttat
     1381 cagcaaccat catgtctgaa tacgccaaca aaaggacacg aggcgctttc atagcagcag
     1441 ttttcgcaat gcaaggcgtg ggaatcatct ttgccggatt ggtttcaatg gtattttcag
     1501 gaatttttaa agcatactac caggctcccc gtttcaatga agacccaatt ttatcaacac
     1561 agcccgaagg ggatttactc tggcgattaa ttctcatgat cggtgctgtt ccagctgcga
     1621 tgacttacta ctggagaatg aaaatgcctg aaaccggtcg ttacactgca atcgtagaag
     1681 ggaacgcgaa acaagccgct gcagacatgg caagagtttt ggacatcgaa atcatagcgg
     1741 agcaagataa gttggctgaa tttaaagcag caaatgatta ccctctttgg tctagcgagt
     1801 ttttcaacag gcacggtcgt catttaatcg gtacaatgag ttgttggttc ttgttagata
     1861 ttgctttcta cagtcagaat ttgacacaga aagatattta ccctgcaatg ggacttattc
     1921 gtcaagacaa agaaatgaat gctattgatg aagttttcca aacttcgcgc gccatgtttg
     1981 ttgtcgcctt gtttggaaca ttccccggtt attggttcac tgttttcttc attgaaaagc
     2041 ttggaagatt caagatacag ttggttggtt tcttcatgat gtctttcttc atgtttgtca
     2101 ttggtgttaa atatgaatac ttgaaagacg aaaacaaaaa cctcttcgct cttttatacg
     2161 ggctaacatt cttcttcgcg aactttgggc ctaatagtac aacttttgtg ttgcccgctg
     2221 agctttttcc tacgcgggtg aggtcaacat gccatgcttt cagtgctgca tctgggaagg
     2281 cgggtgctat ggttggtgcg ttcgggatac aatactacac gttggatggt acgccaagga
     2341 agattagacg cgcaatgatg attctggctt ttacaaattt gataggattc ttttgcacgt
     2401 tcttggtgac ggaaactaag ggacgatctt tggaagagat ttcgggtgag gatggaagag
     2461 agagtgaact cactgcaaca ccaaatgaca gggctccggg gattcgccag gactcaagaa
     2521 ctgagaagat gctcgagccg cggcggccgc cagcttacgt agaacaaaaa ctcatctcag
     2581 aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt tgtagcctta
     2641 gacatgactg ttcctcagtt caagttgggc acttacgaga agaccggtct tgctagattc
     2701 taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact
     2761 tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg
     2821 agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga
     2881 aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga
     2941 ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac
     3001 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     3061 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     3121 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     3181 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     3241 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     3301 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     3361 atagcaatct aatctaaggg 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 ccgacgcggc
     3841 ccgacgggtc cgaggcctcg 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 tttctcatag 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 aagaacagta 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 ca
//
