LOCUS       sequence-437108-        3515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level 0 promoterless vector encoding mutant of 
            Neonothopanus nambi hispidin-3-hydroxylase nnH3H_v2 codon-optimised 
            for expression in Homo sapiens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3515)
  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 3515)
  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: pNK189
FEATURES             Location/Qualifiers
     source          1..3515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      287..875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     776..795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1029..1046
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2428..2446)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2514..2618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2840..3515,1..116)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
       61 tggaagaatt tgtccattac gtaaaaggcg agatcaccaa ggtagtcggc aaataactgt
      121 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      901 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      961 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1021 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     1081 tccccgcgcg ttggccgatt cattaatcac tctgtggtct caaatggctt ctttcgagaa
     1141 cagcctttcc gttcttatag tgggtgcggg cctgggtgga ttggcggctg cgatagcact
     1201 gcgaaggcag ggccacgttg taaaaatata tgaatcatcc tctttcaagg ctgagttggg
     1261 tgccgggctt gcggtgcctc ccaacacctt gaggtcattg caacagcttg ggtgcaatac
     1321 agaaaacctg aatggcgtcg ataatttgtg tttcacggct atgggctacg atggaagtgt
     1381 tgggatgatg aacaatatga ccgactacag ggaggcctat ggcacgtcat ggataatggt
     1441 acatagggtt gatttgcata acgaacttat gagggtggcc cttgatcctg ggggcctcgg
     1501 tccccccgca accctgcatc tcaaccatag ggttactttt tgcgatgtcg atgcctgtac
     1561 cgtaacattt accaacggaa ccactcagtc agccgatttg attgttggtg cagatggaat
     1621 aagatccacg attcggcgct tcgtgctcga agaggacgtc acaatcccag cttctggcat
     1681 agtaggtttt agatggcttg tgcaggctga tgcactcgat ccgtatccag aactcgactg
     1741 gatagtaaag aaacctccac tcggagcaag actcatctct acaccgcaga atccacagag
     1801 tggggtgggg ttggccgacc ggcggacaat tatcatctat gcatgccgag ggggtacgat
     1861 ggtcaatgtt ctcgctgtgc atgacgatga aagagatcag aatacggcag actggtcagt
     1921 gcccgcgagt aaggatgacc tctttagggt gtttcatgat tatcacccac ggttcagacg
     1981 cctgctggaa ttggcccagg acatcaatct ctggcaaatg cgcgtcgtcc cggtcctcaa
     2041 gaaatgggtg aataagcgcg tgtgcctgct cggcgatgca gcgcatgcca tgctccccac
     2101 gctgggccaa ggtttcggga tgggcctcga ggatgccgtg gcgcttggca cgcttttgcc
     2161 aaaggggacc actgccagtc agatcgaaac aagattggct gtttatgaac agctgaggaa
     2221 ggaccgagcg gagtttgtgg cagcagagag ttacgaagaa cagtatgtcc cggagaaacg
     2281 cggactttac ctgagaagca aagaacttag agatcgcgtc atgggatacg atatcaaggt
     2341 ggagtcagag aaagtgttgg aaacgctttt gcgaagctct aatagcgcag gttgagacca
     2401 cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2461 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2521 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2581 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc aactggtcca
     2641 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     2701 actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     2761 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     2821 taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     2881 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     2941 cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc tggttacggt
     3001 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     3061 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     3121 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     3181 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     3241 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     3301 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     3361 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     3421 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     3481 gcgcctgccg gcccagtatc agcccgtcat acttg
//
