LOCUS       sequence-437111-        3050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level 0 promoterless vector encoding mutant of 
            Neonothopanus nambi luciferase nnLuz_v4 codon-optimised for 
            expression in Pichia pastoris, Homo sapiens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3050)
  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 3050)
  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: pNK845
FEATURES             Location/Qualifiers
     source          1..3050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      282..870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     771..790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1024..1041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1958..1976)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2044..2148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2370..3050,1..111)
                     /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 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
       61 gaatttgtcc attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac
      121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
      181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     1081 gcgcgttggc cgattcatta atcactctgt ggtctcaaat gaggtcaact atttccttgt
     1141 cctcccttct tgaaagattg tctaaattga gtagtaggag tatcgctata acttgtggtg
     1201 tcgtacttgc ctctgcaatc gcattcccca taatcaggag agactaccag accttccttg
     1261 aggtgggtcc aagttacgct ccccagaatt tcaggggata cataactgta tgcgtactgt
     1321 cacttttcag acaagaacaa aagggtctgg ccatctacga cagactgcct gaaaaaagaa
     1381 gatggttggc tgatttgccc tttagagaag gaccaagacc atccattact tctcatatca
     1441 ttcaaaggca aaggactcaa ttggtagacc aggaatttgc aactagggag ctgattgata
     1501 aggtgatacc cagagtacaa gctagacata cagacaagac tttcctttct acatctaaat
     1561 tcgagttcca tgctaaggcc atcttcctgc ttcctagtat tccaattaac gaccccttga
     1621 acattccatc ccatgatact gtcaggagga caaagagaga aatcgcccat atgcacgact
     1681 accatgattg ttctttgcac cttgcactgg caccacagga cggcaaggag gtcctgaaaa
     1741 agggctgggg tcaaagacac cccctggcag gtcctggtgt acccggacca cctactgaat
     1801 ggacattctt gtatgctcca aggaacgagg aggaggcaag ggtggtggag atgatcgtag
     1861 aagcaagtat tggatatatg acgaacgatc ctgctggtaa aattgtggaa aatgctaaag
     1921 gttgagacca cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta
     1981 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2041 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2101 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc
     2161 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     2221 gcttgttatg actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     2281 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     2341 gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga
     2401 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     2461 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc
     2521 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     2581 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     2641 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     2701 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     2761 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     2821 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     2881 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     2941 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     3001 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
//
