LOCUS       sequence-231086-        4774 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 4774)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4774)
  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: NiaP
FEATURES             Location/Qualifiers
     source          1..4774
                     /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             2443..2466
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2467..2490
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2500..2517
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2597..2843
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2858..3271
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3278..3325
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3344..3718
                     /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      3784..4031
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3800..3818)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4106..4694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4186..4205)
                     /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 ggctcctgaa
      961 atgatggagg aggggcaatc agcttcctat accgtggatg acgctctctt gtcatcaggt
     1021 tttgggaggt tccagatatt gattctttca tatgctggga ttggcttaat tgccgaagcg
     1081 atggaaatga tgctgctatc atttgttggt ccatcagtac agctagaatg gaagcttact
     1141 tctcatcagg aaagcatgat tacaagtgtt gtttttgttg ggatgctgat tggagcttac
     1201 tcgtggggtg tggtttcaga caactatgga aggaggagag gctttctctt tacagccatt
     1261 atgacaagtg gagctggatt ttttagcgct tttgctccga actatctatc attaatttcc
     1321 ctgagattct tagttggtat tggtttggga ggaggacctg ttctgggatc ttggttcttg
     1381 gaatttgttc ctgctccaac tagaggaact tggatggtgg tattttctgc attttggact
     1441 gttggaacaa tcttggaggc atctcttgca tggacaatta tgcccaagtt tggttggagg
     1501 tggttgttag cattatcagc tgttccatct tttctcctgc tgctatttta tgccattaca
     1561 ccagagtcac caaggtttct ttgcatgaaa ggccggacta ctgaggctgt ggatatattg
     1621 gagaaaatgg caagactaaa caatgtacaa ctaccttctg gtaggcttgt ttctgacaaa
     1681 aatattgagc tagatgaagt ttcaggatct tcagagtcca cgacacttct ggctggtgcc
     1741 gaagaaagtg acaacctaaa tgaggatcag ggttctgatt tcggaggtat taagtccgtc
     1801 ggcaagctac tcgcaccaaa actgatcaga gcaaccctgc ttctgtggat ggccttcttt
     1861 ggaaatgcgt ttgcctatta tggcattgtt cttctgacat cagagctcag taatggaaac
     1921 aggatatgtg ccaaagagga tgttgaatca gttcattcaa ccaatgcaag cctatataag
     1981 aatgttttca tatctagttt tgcagagatc ccagggtcat ttctgtcggc catgatcgtg
     2041 gatagatttg gtcggaagcg ttccatggcg tcgatgctct tcactagctg cgttttctta
     2101 ctcccactag tgttctcccg gacagatata ctaaccagaa tctccctgtt tggtgcccgg
     2161 ctctgtatat ctgccagctt caccatcgta tacatatatg ctccagagat ttacccgacc
     2221 gcggtgagga cgacgggcat cggcatcgcg agctcagtgg gcaggatcgg gggcatcctc
     2281 tgcccactcg tcgctgtcgc cctggtgcac agctgccagc agacgacggc gatcctcctc
     2341 ttcgagcttg taatcttcct ctccggcctg gccgtctcgt tcttcccctt cgagacgaag
     2401 gggtgtaggc tcaacgacac cgacgtggac atgaacggta ccttggaagt tttgttccag
     2461 ggtccacatc atcaccacca tcaccatcat tgagtcgacc atcatcatca tcatcattga
     2521 gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg
     2581 tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt
     2641 catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt
     2701 ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg
     2761 taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc
     2821 agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc
     2881 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     2941 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     3001 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
     3061 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     3121 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa
     3181 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     3241 ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata
     3301 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     3361 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     3421 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     3481 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     3541 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     3601 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3661 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3721 cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg
     3781 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     3841 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     3901 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     3961 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     4021 ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4741 aaaactcacg ttaagggatt ttggtcatga gatc
//
