LOCUS       sequence-231203-        4922 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 4922)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4922)
  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: OsNRAT-uPIC
FEATURES             Location/Qualifiers
     source          1..4922
                     /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             2621..2641
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2648..2665
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2745..2991
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3006..3419
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3426..3473
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3492..3866
                     /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      3932..4179
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3948..3966)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4334..4353)
                     /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 tatggaaggg actggtgaga tgagagaggt tggacgagaa actctccatg gaggtgtggt
     1021 gcagagtgtt agtgagactg atgagtacaa agaaaagact attgacagtg aaaaagatgg
     1081 gcagtttcgg gtgcagccaa gatggaggaa attcttggct catgttgggc ctggagctct
     1141 ggtggccatt ggcttcctag atcctagcaa cttggaaact gacatgcaag ctggagctga
     1201 cttcaagtat gagcttctgt gggtcatctt agttgggatg gtctttgcac ttctgatcca
     1261 aacattagct gccaaccttg gagtgaaaac aggaaggcac cttgctgaac tgtgcaggga
     1321 agagtacccg cactatgtca acattttctt atggatcatt gcggagttgg cggtgatttc
     1381 tgatgacatt cctgaagtgt tgggcacagc ttttgcgttc aacatattgc ttaagattcc
     1441 agtgtgggct ggagttattc tcacagtgtt cagcaccctc ttgcttcttg gagtacagag
     1501 atttggggct cggaagctgg agttcatcat agcagcattc atgttcacta tggcagcttg
     1561 cttcttcgga gagctaagct atttgaggcc gtctgcagga gaggtggtca agggcatgtt
     1621 tgtcccttct ctccaaggga aaggcgctgc tgccaatgcg atcgcactct ttggcgctat
     1681 catcacacca tacaacttgt tcttgcactc tgctctggtc ctgtcaagga agaccccacg
     1741 atcagacaaa agcatcagag ctgcttgcag atacttcctt attgagtgta gtcttgcatt
     1801 catcgtggct ttcctcataa atgtttcagt cgttgttgtt gctggatcca tttgcaatgc
     1861 gaataatctt tctccagctg atgccaacac gtgtggtgac ctaactctgc aatccacacc
     1921 tcttttactc aggaatgttt tggggagatc aagctcggtg gtgtatgctg ttgcgctgct
     1981 agcttctggg caaagcacca ctatcagttg cacgttcgct gggcaggtca tcatgcaggg
     2041 gtttctggac atgaagatga agaattgggt aaggaacctc atcacccgag tcatcgccat
     2101 agctcctagc ctcatcgtct ccattgtcag tggtccctca ggcgccggca agctaatcat
     2161 cctgtcatca atgatcctgt catttgagct gccgttcgct ctcatccctc tactcaagtt
     2221 ctgcaacagc agcaagaaag ttggacccct caaggaatcc atctatacgg tggtgatcgc
     2281 gtggatcctg agcttcgcgc tcatcgtcgt caacacctac ttcctagtgt ggacatacgt
     2341 ggactggctc gtccacaaca acctccccaa gtatgccaat ggcctcatct ccgtggtggt
     2401 gttcgcgctc atggccgcct acctggtcgc cgtcgtctac ctgacgttca ggaaggacac
     2461 ggtggcgacg tacgtgccgg tgccggagcg ggcgcaggcg caggtggagg ccggcgggac
     2521 gccggtggtg gacgcctccg ccgccgacga ggaccagccg gctccgtaca ggaaggacct
     2581 tgccgatgct tccatggagg cattaaacct cagcagcggc gagaacctgt acttccaatc
     2641 tgtcgaccat catcatcatc atcattgagt ttgtagcctt agacatgact gttcctcagt
     2701 tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga
     2761 atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata
     2821 tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc
     2881 tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg
     2941 tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt
     3001 gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag
     3061 attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa
     3121 atttcccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag
     3181 aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca
     3241 cgtttctttt tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt
     3301 gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc
     3361 tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagg
     3421 gcggtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3481 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3541 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3601 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3661 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     3721 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     3781 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     3841 acttcgtggc cgaggagcag gactgacacg tccgacgcgg cccgacgggt ccgaggcctc
     3901 ggagatccgt cccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     3961 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     4021 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     4081 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     4141 cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctggagac caacatgtga
     4201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4681 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4921 tc
//
