LOCUS       sequence-438219-        5744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression Pv4CL1 in yeast (P.pastoris).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5744)
  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 5744)
  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: pNK7188
FEATURES             Location/Qualifiers
     source          1..5744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1007..1483
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             3195..3224
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3240..3257
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3337..3582
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3598..4011
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        4018..4065
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4122..5147
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTCYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     terminator      5246..5493
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5262..5280)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(join(5562..5744,1..406))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5642..5661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
       61 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      481 gagatcagat ctctgctact ctggtcccaa gtgaaccacc ttttggaccc tattgaccgg
      541 accttaactt gccaaaccta aacgcttaat gcctcagacg ttttaatgcc tctcaacacc
      601 tccaaggttg ctttcttgag catgcctact aggaacttta acgaactgtg gggttgtaga
      661 cagtttcagg cgtgtcccga ccaatatggc ctactagact ctctgaaaaa tcacagtttt
      721 ccagtagttc cgatcaaatt accatcgaaa tggtcccata aacggacatt tgacatccgt
      781 tcctgaatta tagtcttcca ccgtggatca tggtgttcct ttttttccca aagaatatca
      841 gcatccctta actacgttag gtcagtgatg acaatggacc aaattgttgc aaggtttttc
      901 tttttctttc atcggcacat ttcagcctca catgcgacta ttatcgatca atgaaatcca
      961 tcaagattga aatcttaaaa ttgccccttt cacttgacag gatccttttt tgtagaaatg
     1021 tcttggtgtc ctcgtccaat caggtagcca tctctgaaat atctggctcc gttgcaactc
     1081 cgaacgacct gctggcaacg taaaattctc cggggtaaaa cttaaatgtg gagtaatgga
     1141 accagaaacg tctcttccct tctctctcct tccaccgccc gttaccgtcc ctaggaaatt
     1201 ttactctgct ggagagcttc ttctacggcc cccttgcagc aatgctcttc ccagcattac
     1261 gttgcgggta aaacggaggt cgtgtacccg acctagcagc ccagggatgg aaaagtcccg
     1321 gccgtcgctg gcaataatag cgggcggacg catgtcatga gattattgga aaccaccaga
     1381 atcgaatata aaaggcgaac acctttccca attttggttt ctcctgaccc aaagacttta
     1441 aatttaattt atttgtccct atttcaatca attgaacaac tatcaattcg aagaggaatt
     1501 caagcttcat atggctagca aatggggtca gttgatgagt cagcagcagc agaagtggtc
     1561 tttaggtcaa agcttccaga tatcgaaata aataattcac aacctttgca cgcttattgc
     1621 tttggcaaga tgccagaagt ggccggtaga ccatgcctta tagacggtca aacaggtgct
     1681 agttatacct acgcagaggt cgaaagtctt agtaggagag cagctgccgg acttaggagg
     1741 atgggtgtgg gaaaaggtga cgtcgtcatg tcattgctta gaaattgtcc tgaattcgcc
     1801 ttcgcttttc ttggagctgc cagacttggt gccgctacta ccactgcaaa ccctttttac
     1861 actcctcacg aaatacacag acaagctgag gcagccggtg ctaaattgat agtgactgag
     1921 gcttgcgccg tggataaggt gagggaattc gccgtagcta ggggtatccc agttgttaca
     1981 gttgacggca gattcgatgg ttgcgctgag ttcgctgaag tcattgctgc agaggagctt
     2041 gaggccgatg cagacataca gcctgatgat gtagtcgctt tgccatactc tagtgggaca
     2101 actggattgc caaaaggagt aatgcttact catagatctc ttatcacatc tgttgctcaa
     2161 caggtggacg gtgagaatcc taatttgcac ttcaggaagg acgacgtttt gctttgcttg
     2221 ttgcctttgt ttcacatcta ctcattgaat agtgtattgc ttgccggctt gagagcaggg
     2281 tgtgcaattg tgattatgag aaaattcgaa actggggcac ttgtagaatt ggtgagggct
     2341 catggagtaa ctgtcgcccc ttttgtacct ccaattgtgg tagaaattgc caagtctcct
     2401 agggttgggg ccgctgattt ggcttcaatc agaatggtaa tgtcaggggc cgctcctatg
     2461 ggaaaagact tgcaagacgc tttcatggcc aagattccta atgcagtgtt gggtcagggc
     2521 tacggtatga cagaggccgg accagtgttg gcaatgtgcc ttgcctttgc aaaggagcca
     2581 ttccaggtca aatcagggtc atgcggcact gtggtcagaa acgcagaatt gaagatagtt
     2641 gatcctgata ccggtgccgc acttggaaga aatcagccag gcgaaatatg tatcagaggt
     2701 gagcagataa tgaaggggta cttgaacgat cctgaatcta ccaaaaatac aatagataag
     2761 gatggttggc ttcacaccgg agacataggc tacgtggacg acgacgacga aatattcatt
     2821 gttgacaggt tgaaggaaat catcaaatac aaaggatttc aagtcccacc agccgaattg
     2881 gaggctttgt tgatcactca tccagaaata aaagatgcag ccgttgtcag tatgaaagac
     2941 gatttggccg gtgaaatccc agtggctttc atcgtcagga ctgaggggag tcagttgaca
     3001 gaggacgaaa taaaacaatt tgttgctaaa gaagttgtat tctataaaaa gatacacaag
     3061 gtctttttta ctgattctat acctaagaat ccatcaggga agattttgag gaaagatctt
     3121 agggctagac ttgccgcagg ggtacattca ggtagttaac tagcttagct aaataagcgg
     3181 ccgccagctt tctagaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc
     3241 atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg
     3301 ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt
     3361 tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag
     3421 gatttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc ctatctcgca
     3481 gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat gtttttcttg
     3541 gtatttccca ctcctcttca gagtacagaa gattaagtga gaaccttcgt ttgtgcggat
     3601 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
     3661 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc
     3721 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
     3781 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
     3841 tttcttgaaa attttttttt ttgatttttt tctctttcga cgacctccca ttgatattta
     3901 agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa
     3961 ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt
     4021 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4081 accatggatg tctctattca acgggaaacg tcttgctcga gatgaaaaag cctgaactca
     4141 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     4201 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     4261 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     4321 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     4381 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     4441 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     4501 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4561 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4621 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4681 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     4741 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     4801 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgt
     4861 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     4921 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     4981 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     5041 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     5101 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataaact agtcacttcg
     5161 tggccgagga gcaggactga cacgtccgac gcggcccgac gggtccgagg cctcggagat
     5221 ccgtccccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     5281 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     5341 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     5401 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     5461 aaggttttgg gacgctcgaa ggctttaatt tgcaagctga acatgtgagc aaaaggccag
     5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgt
//
