LOCUS       sequence-437109-        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 Pichia pastoris.
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: pNK4169
FEATURES             Location/Qualifiers
     source          1..3515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      284..872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     773..792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1026..1043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2425..2443)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2511..2615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2837..3515,1..113)
                     /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 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
       61 aagaatttgt ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag
      121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
      181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
      241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      421 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      901 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      961 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     1021 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     1081 ccgcgcgttg gccgattcat taatcactct gtggtctcaa atggcctcat tcgagaattc
     1141 attgtcagtg cttattgttg gtgctggtct tggtggtttg gcagctgcaa tagcacttag
     1201 gaggcagggt catgtggtca aaatatatga atcttcatca ttcaaagcag agttgggagc
     1261 aggtcttgcc gttcctccaa atacccttag gtcattgcaa caattgggat gtaatacaga
     1321 gaatttgaat ggagtcgata acttgtgctt caccgctatg ggatatgacg gttctgtcgg
     1381 tatgatgaac aacatgacag attacagaga ggcttatggt acatcatgga tcatggtgca
     1441 cagagtcgat ttgcacaatg aacttatgag ggttgccttg gatccaggag gtttgggacc
     1501 ccctgctaca ttgcacttga accacagggt caccttttgt gatgttgacg cttgcaccgt
     1561 tacattcaca aatggtacaa cccaatcagc tgatttgatc gtgggagccg atggaataag
     1621 gtcaacaatt agaagatttg tgcttgaaga ggacgtcaca atccctgcct caggtatcgt
     1681 tggttttagg tggcttgtcc aagccgatgc tcttgatcct tacccagaat tggattggat
     1741 agttaagaaa ccaccacttg gagccagact tatatctacc cctcagaacc cacagtctgg
     1801 tgttggtttg gctgacagga gaactattat aatatatgca tgtaggggag gtactatggt
     1861 taacgttctt gctgttcacg atgatgagag agatcagaac acagcagatt ggtcagtgcc
     1921 agcatcaaaa gatgaccttt ttagggtgtt ccatgactat cacccgaggt tcaggaggct
     1981 tttggagttg gcacaggaca taaatctttg gcaaatgagg gttgtgccag tgttgaaaaa
     2041 atgggttaac aagagggtgt gccttcttgg cgacgcagca catgcaatgt tgcctacatt
     2101 gggtcaaggt ttcggaatgg gattggagga cgcagtcgcc cttggtacac ttcttccaaa
     2161 gggtacaact gcatctcaga ttgaaaccag gcttgctgtg tacgaacaat tgaggaaaga
     2221 tagggcagag tttgtggctg ccgagtctta cgaagaacaa tacgtgcctg aaaaaagagg
     2281 tctttatctt aggtctaagg aacttaggga tagagttatg ggttacgaca tcaaggttga
     2341 gtctgaaaag gtgttggaaa ccttgttgcg ttcttcaaac tcagcaggtt gagaccacga
     2401 agtggctctt cagtggacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2461 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtatgcg ctcacgcaac tggtccagaa
     2641 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2701 gttttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     2761 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     2821 aacaaagtta aacatcatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     2881 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     2941 ctccgcagtg gatggcggcc tgaagccaca cagcgatatt gatttgctgg ttacggtgac
     3001 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     3061 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     3121 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     3181 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     3241 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     3301 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     3361 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     3421 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     3481 cctgccggcc cagtatcagc ccgtcatact tgaag
//
