LOCUS       sequence-438291-        4646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression PpASCL in yeast (P.pastoris).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4646)
  AUTHORS   Palkina KA, Karataeva TA, Perfilov MM, Fakhranurova LI, Markina NM, 
            Somermeyer LG, Garcia-Perez E, Vazquez-Vilar M, Rodriguez-Rodriguez 
            M, Vazquez-Vilriales V, Shakhova ES, Mitiouchkina T, Belozerova OA, 
            Kovalchuk SI, Alekberova A, Malyshevskaia AK, Bugaeva EN, Guglya EB,
            Balakireva A, Sytov N, Bezlikhotnova A, Boldyreva DI, Babenko VV, 
            Kondrashov FA, Choob VV, Orzaez D, Yampolsky IV, Mishin AS, 
            Sarkisyan KS
  TITLE     A hybrid pathway for self-sustained luminescence.
  JOURNAL   Sci Adv. 2024 Mar 8;10(10):eadk1992. doi: 10.1126/sciadv.adk1992. 
            Epub 2024 Mar 8.
  PUBMED    38457503
REFERENCE   2  (bases 1 to 4646)
  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"; journalName: "Sci Adv."; 
            date: "2024-03-8"; volume: "10(10):eadk1992. doi"; pages: " 
            10.1126/sciadv.adk1992. Epub 2024 Mar 8"
COMMENT     Sequence Label: pNK7182
FEATURES             Location/Qualifiers
     source          1..4646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        452..928
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2195..2212
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2258..2287
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2303..2320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2400..2646
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2661..3074
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3081..3128
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3147..3521
                     /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      3587..3834
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3603..3621)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctaaacgc ttaatgcctc agacgtttta atgcctctca acacctccaa ggttgctttc
       61 ttgagcatgc ctactaggaa ctttaacgaa ctgtggggtt gcagacagtt tcaggcgtgt
      121 cccgaccaat atggcctact agactctctg aaaaatcaca gttttccagt agttccgatc
      181 aaattaccat cgaaatggtc ccataaacgg acatttgaca tccgttcctg aattatagtc
      241 ttccaccgtg gatcatggtg ttcctttttt tcccaaagaa tatcagcatc ccttaactac
      301 gttaggtcag tgatgacaat ggaccaaatt gttgcaaggt ttttcttttt ctttcatcgg
      361 cacatttcag cctcacatgc gactattatc gatcaatgaa atccatcaag attgaaatct
      421 taaaattgcc cctttcactt gacaggatcc ttttttgtag aaatgtcttg gtgtcctcgt
      481 ccaatcaggt agccatctct gaaatatctg gctccgttgc aactccgaac gacctgctgg
      541 caacgtaaaa ttctccgggg taaaacttaa atgtggagta atggaaccag aaacgtctct
      601 tcccttctct ctccttccac cgcccgttac cgtccctagg aaattttact ctgctggaga
      661 gcttcttcta cggccccctt gcagcaatgc tcttcccagc attacgttgc gggtaaaacg
      721 gaggtcgtgt acccgaccta gcagcccagg gatggaaaag tcccggccgt cgctggcaat
      781 aatagcgggc ggacgcatgt catgagatta ttggaaacca ccagaatcga atataaaagg
      841 cgaacacctt tcccaatttt ggtttctcct gacccaaaga ctttaaattt aatttatttg
      901 tccctatttc aatcaattga acaactatca attcgaaacg atggctagtc gtcgtgtcga
      961 ggcagcattc gatggacaag cagtagaact gggtgctact attcctgccg ccaatggcaa
     1021 cggaactcat cagtcaatta aggttcctgg acatcgtcaa gtgactccag gcaagactac
     1081 gattatggct attggccgtg ctgtgcccgc taacacgacg ttcaatgatg gtttggcaga
     1141 ccattacatc caggagttca atctgcaaga tccagtcctt caggctaaac tgcgtcgttt
     1201 gtgcgaaacc acaaccgtga aaacgcgtta tctggtcgtc aataaggaga ttctggatga
     1261 gcatcctgag ttcctggttg atggagctgc aaccgtatct cagcgtctgg ctattacagg
     1321 cgaagctgtt acccaacttg gacatgaggc agcaacggct gccatcaaag agtggggccg
     1381 tccagcctcc gaaattaccc atcttgttta tgtttcctca tccgaaatcc gtctgccagg
     1441 tggcgacttg tacctggcac aattgcttgg cttgcgttca gacgtgaacc gtgtaatgtt
     1501 gtacatgctg ggttgctatg gtggtgcatc tggaatccgt gttgccaaag acttggctga
     1561 gaataaccca ggctcacgtg tacttctgat cacgtcagaa tgcactttga ttggatataa
     1621 gtcactttcc cctgaccgtc cttacgacct ggttggagca gctctgttcg gtgacggtgc
     1681 tgccgctatg attatgggaa aagatccaat ccctgttttg gagcgtgctt tctttgagct
     1741 tgactgggcc ggtcagtctt ttattcctgg aactaacaaa accatcgatg gtcgtttgtc
     1801 tgaagaaggc atttccttca agctgggtcg tgaactgcct aagttgattg aatccaatat
     1861 tcagggtttc tgcgatccta tcttgaaacg tgctggtggt ctgaagtata acgatatttt
     1921 ctgggcagtt cacccaggcg gaccagctat cttgaatgct gtccagaaac aactggattt
     1981 ggcacccgag aaacttcaga ccgcacgtca ggttctgcgt gactacggta acatttctag
     2041 ttccacctgc atttacgtac tggactatat gcgtcatcaa tccttgaaac tgaaggaagc
     2101 aaatgacaac gtaaatacgg aaccagagtg gggtctgctg cttgcatttg gccccggcgt
     2161 cactatcgag ggtgctttgc tgcgtaacct gtgccatcac caccaccatc actaaactag
     2221 ttcggtacct cgagccgcgg cggccgccag ctttctagaa caaaaactca tctcagaaga
     2281 ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttgta gccttagaca
     2341 tgactgttcc tcagttcaag ttgggcactt acgagaagac cggtcttgct agattctaat
     2401 caagaggatg tcagaatgcc atttgcctga gagatgcagg cttcattttt gatacttttt
     2461 tatttgtaac ctatatagta taggattttt tttgtcattt tgtttcttct cgtacgagct
     2521 tgctcctgat cagcctatct cgcagctgat gaatatcttg tggtaggggt ttgggaaaat
     2581 cattcgagtt tgatgttttt cttggtattt cccactcctc ttcagagtac agaagattaa
     2641 gtgagacctt cgtttgtgcg gatcccccac acaccatagc ttcaaaatgt ttctactcct
     2701 tttttactct tccagatttt ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca
     2761 agcacagcat actaaatttc ccctctttct tcctctaggg tgtcgttaat tacccgtact
     2821 aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa
     2881 taaaaatttt tatcacgttt ctttttcttg aaaatttttt tttttgattt ttttctcttt
     2941 cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt
     3001 tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag
     3061 caatctaatc taagggcggt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3121 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     3181 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     3241 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     3301 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     3361 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     3421 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     3481 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtccga cgcggcccga
     3541 cgggtccgag gcctcggaga tccgtccccc ttttcctttg tcgatatcat gtaattagtt
     3601 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     3661 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     3721 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     3781 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctg
     3841 gagaccaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagatcaga tctctgctac tctggtccca agtgaaccac cttttggacc
     4621 ctattgaccg gaccttaact tgccaa
//
