LOCUS       sequence-437148-        4674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGAP-like vector encoding mutant of Neonothopanus nambi luciferase 
            nnLuz_v4 under control of GAP promoter, for yeast expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4674)
  AUTHORS   Shakhova ES, Karataeva TA, Markina NM, Mitiouchkina T, Palkina KA, 
            Perfilov MM, Wood MG, Hoang TT, Hall MP, Fakhranurova LI, Alekberova
            AE, Malyshevskaia AK, Gorbachev DA, Bugaeva EN, Pletneva LK, Babenko
            VV, Boldyreva DI, Gorokhovatsky AY, Balakireva AV, Gao F, Choob VV, 
            Encell LP, Wood KV, Yampolsky IV, Sarkisyan KS, Mishin AS
  TITLE     An improved pathway for autonomous bioluminescence imaging in 
            eukaryotes.
  JOURNAL   Nat Methods. 2024 Mar;21(3):406-410. doi: 
            10.1038/s41592-023-02152-y. Epub 2024 Jan 22.
  PUBMED    38253843
REFERENCE   2  (bases 1 to 4674)
  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"; doi: 
            "10.1038/s41592-023-02152-y"; journalName: "Nat Methods"; date: 
            "2024-03"; volume: "21"; issue: "3"; pages: "406-410"
COMMENT     Sequence Label: pNK5785
FEATURES             Location/Qualifiers
     source          1..4674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             852..881
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             897..914
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      994..1239
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1255..1668
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1675..1722
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(1817..1836)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2658..2905
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2674..2692)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2974..3562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3054..3073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4163..4639
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
ORIGIN
        1 caaaaaatga ggtcaactat ttccttgtcc tcccttcttg aaagattgtc taaattgagt
       61 agtaggagta tcgctataac ttgtggtgtc gtacttgcct ctgcaatcgc attccccata
      121 atcaggagag actaccagac cttccttgag gtgggtccaa gttacgctcc ccagaatttc
      181 aggggataca taactgtatg cgtactgtca cttttcagac aagaacaaaa gggtctggcc
      241 atctacgaca gactgcctga aaaaagaaga tggttggctg atttgccctt tagagaagga
      301 ccaagaccat ccattacttc tcatatcatt caaaggcaaa ggactcaatt ggtagaccag
      361 gaatttgcaa ctagggagct gattgataag gtgataccca gagtacaagc tagacataca
      421 gacaagactt tcctttctac atctaaattc gagttccatg ctaaggccat cttcctgctt
      481 cctagtattc caattaacga ccccttgaac attccatccc atgatactgt caggaggaca
      541 aagagagaaa tcgcccatat gcacgactac catgattgtt ctttgcacct tgcactggca
      601 ccacaggacg gcaaggaggt cctgaaaaag ggctggggtc aaagacaccc cctggcaggt
      661 cctggtgtac ccggaccacc tactgaatgg acattcttgt atgctccaag gaacgaggag
      721 gaggcaaggg tggtggagat gatcgtagaa gcaagtattg gatatatgac gaacgatcct
      781 gctggtaaaa ttgtggaaaa tgctaaaggt agttaactag cttagctaaa taagcggccg
      841 ccagctttct agaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc
      901 atcatcatca tcattgagtt tgtagcctta gacatgactg ttcctcagtt caagttgggc
      961 acttacgaga agaccggtct tgctagattc taatcaagag gatgtcagaa tgccatttgc
     1021 ctgagagatg caggcttcat ttttgatact tttttatttg taacctatat agtataggat
     1081 ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct
     1141 gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta
     1201 tttcccactc ctcttcagag tacagaagat taagtgagaa ccttcgtttg tgcggatccc
     1261 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga
     1321 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct
     1381 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac
     1441 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt
     1501 cttgaaaatt tttttttttg atttttttct ctttcgacga cctcccattg atatttaagt
     1561 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
     1621 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaaggg cggtgttgac
     1681 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     1741 atggatgtct catattcacg ggaaacgtct tgctcgaggc cgcgattaaa ttccaacatg
     1801 gatgctgatt tatgtgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1861 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1921 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1981 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2041 gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2101 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2161 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2221 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2281 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2341 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2401 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2461 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2521 aaattgcagt ttcatttgat gctcgatgag tttttctaaa ctagtcactt cgtggccgag
     2581 gagcaggact gacacgtccg acgcggcccg acgggtccga ggcctcggag atccgtcccc
     2641 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     2701 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     2761 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     2821 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     2881 gggacgctcg aaggctttaa tttgcaagct gaacatgtga gcaaaaggcc agcaaaaggc
     2941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga tcagatctct gctactctgg
     3661 tcccaagtga accacctttt ggaccctatt gaccggacct taacttgcca aacctaaacg
     3721 cttaatgcct cagacgtttt aatgcctctc aacacctcca aggttgcttt cttgagcatg
     3781 cctactagga actttaacga actgtggggt tgtagacagt ttcaggcgtg tcccgaccaa
     3841 tatggcctac tagactctct gaaaaatcac agttttccag tagttccgat caaattacca
     3901 tcgaaatggt cccataaacg gacatttgac atccgttcct gaattatagt cttccaccgt
     3961 ggatcatggt gttccttttt ttcccaaaga atatcagcat cccttaacta cgttaggtca
     4021 gtgatgacaa tggaccaaat tgttgcaagg tttttctttt tctttcatcg gcacatttca
     4081 gcctcacatg cgactattat cgatcaatga aatccatcaa gattgaaatc ttaaaattgc
     4141 ccctttcact tgacaggatc cttttttgta gaaatgtctt ggtgtcctcg tccaatcagg
     4201 tagccatctc tgaaatatct ggctccgttg caactccgaa cgacctgctg gcaacgtaaa
     4261 attctccggg gtaaaactta aatgtggagt aatggaacca gaaacgtctc ttcccttctc
     4321 tctccttcca ccgcccgtta ccgtccctag gaaattttac tctgctggag agcttcttct
     4381 acggccccct tgcagcaatg ctcttcccag cattacgttg cgggtaaaac ggaggtcgtg
     4441 tacccgacct agcagcccag ggatggaaaa gtcccggccg tcgctggcaa taatagcggg
     4501 cggacgcatg tcatgagatt attggaaacc accagaatcg aatataaaag gcgaacacct
     4561 ttcccaattt tggtttctcc tgacccaaag actttaaatt taatttattt gtccctattt
     4621 caatcaattg aacaactatc aattcgaaga ggaattcaag cttcatatgg ctag
//
