LOCUS       sequence-225002-        4273 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 4273)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4273)
  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: AtTPT (ORF-HIS)
FEATURES             Location/Qualifiers
     source          1..4273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1002..1025
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1026..1049
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1059..1076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1156..1402
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1417..1830
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1837..1884
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1903..2277
                     /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      2343..2590
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2359..2377)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2665..3253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2745..2764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3335..4273
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 aggaattcat ggccgccgct gctgaaggtg gagataccgc tggggatgct aaagttggat
       61 tcctcgccaa gtatccatgg ctagtcactg gattcttctt cttcatgtgg tacttcttga
      121 atgtgatttt caacatcctt aacaagaaga tctataatta cttcccttat ccctattttg
      181 tatcggtgat acacttgttc gtgggagttg tttactgctt gatcagctgg tccgtgggtc
      241 ttcctaaacg tgccccgatt gactcgaacc tcctcaaggt attgatacca gtcgcagtct
      301 gtcacgcctt aggccatgtc actagcaatg tctctttcgc tgcggttgct gtctccttca
      361 ctcacaccat caaagcactt gagccattct tcaatgcggc tgcttctcaa ttcattatgg
      421 gacaatccat ccccataaca ctatggttgt ctctagctcc tgttgttctt ggtgttgcaa
      481 tggcttcact aactgagcta tcattcaact ggctcggttt catcagtgct atgatatcaa
      541 acatttcttt cacttaccga agcatcttct ccaagaaagc catgactgat atggacagta
      601 caaatgtcta cgcttacatc tccatcatcg cactcttcgt ctgcattcct cctgccatca
      661 tcgttgaagg tcctaaacta cttaaccatg gtttcgccga cgcgattgct aaagttggaa
      721 tgactaaatt catctctgat ctcttctggg ttggaatgtt ttaccatctc tacaatcagc
      781 tggctaccaa taccttggag agggttgcac cgctgactca cgcggttgga aacgttctga
      841 aacgtgtgtt cgtgatcggt ttctccatcg ttatcttcgg aaacaagata tcgacacaga
      901 caggtatagg aacaggaata gccattgctg gagttgcaat gtactctatc attaaggcca
      961 agatcgaaga agagaaacgg caaggaaaga aagcaggtac cttggaggtt ttgttccaag
     1021 gtccacatca tcaccaccat caccatcact gagtcgacca tcatcatcat catcattgag
     1081 tttgtagcct tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt
     1141 cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc
     1201 atttttgata cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt
     1261 tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt
     1321 aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca
     1381 gagtacagaa gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca
     1441 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
     1501 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
     1561 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
     1621 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
     1681 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
     1741 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
     1801 tagaaagaaa gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag
     1861 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     1921 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     1981 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     2041 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     2101 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     2161 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     2221 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     2281 gtccgacgcg gcccgacggg tccgaggcct cggagatccg tccccctttt cctttgtcga
     2341 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     2401 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     2461 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     2521 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     2581 tttaatttgc aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3301 aaactcacgt taagggattt tggtcatgag atcagatcta acatccaaag acgaaaggtt
     3361 gaatgaaacc tttttgccat ccgacatcca caggtccatt ctcacacata agtgccaaac
     3421 gcaacaggag gggatacact agcagcagac cgttgcaaac gcaggacctc cactcctctt
     3481 ctcctcaaca cccacttttg ccatcgaaaa accagcccag ttattgggct tgattggagc
     3541 tcgctcattc caattccttc tattaggcta ctaacaccat gactttatta gcctgtctat
     3601 cctggccccc ctggcgaggt tcatgtttgt ttatttccga atgcaacaag ctccgcatta
     3661 cacccgaaca tcactccaga tgagggcttt ctgagtgtgg ggtcaaatag tttcatgttc
     3721 cccaaatggc ccaaaactga cagtttaaac gctgtcttgg aacctaatat gacaaaagcg
     3781 tgatctcatc caagatgaac taagtttggt tcgttgaaat gctaacggcc agttggtcaa
     3841 aaagaaactt ccaaaagtcg gcataccgtt tgtcttgttt ggtattgatt gacgaatgct
     3901 caaaaataat ctcattaatg cttagcgcag tctctctatc gcttctgaac cccggtgcac
     3961 ctgtgccgaa acgcaaatgg ggaaacaccc gctttttgga tgattatgca ttgtctccac
     4021 attgtatgct tccaagattc tggtgggaat actgctgata gcctaacgtt catgatcaaa
     4081 atttaactgt tctaacccct acttgacagc aatatataaa cagaaggaag ctgccctgtc
     4141 ttaaaccttt ttttttatca tcattattag cttactttca taattgcgac tggttccaat
     4201 tgacaagctt ttgattttaa cgacttttaa cgacaacttg agaagatcaa aaaacaacta
     4261 attattcgaa acg
//
