LOCUS       sequence-450697-        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 Nicotiana benthamiana.
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: pNK115
FEATURES             Location/Qualifiers
     source          1..3050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     230..249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     483..500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1048..1059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1417..1435)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1503..1607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1829..2620
                     /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"
     rep_origin      join(2791..3050,1..329)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
       61 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      121 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      181 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      241 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      301 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      361 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      421 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      481 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
      541 cgcgttggcc gattcattaa tcactctgtg gtctcaaatg aggtctacta tctctttgtc
      601 tagtctcctt gagagactga gcaaattgtc atcccgatcc atcgcaatta cctgcggcgt
      661 agtcttggca agtgcaatag cattcccaat tatccgaaga gactatcaaa cgttccttga
      721 agtcggtccg agctatgccc cacagaactt ccgaggctat atcactgttt gtgtcttgtc
      781 actttttagg caagagcaga aagggttggc aatctatgac aggcttccag aaaagaggcg
      841 ttggcttgcc gacttgccgt ttcgtgaagg gccaaggcca tccataacct cccacattat
      901 acagcgtcag cgtactcagc tggtagatca agagtttgca accagagaac ttatcgataa
      961 ggtgatccca cgtgtgcaag caagacacac agacaaaact tttctcagta cgtcaaaatt
     1021 tgagtttcat gcaaaggcca tcttccttct cccctctatc cctattaatg atcctcttaa
     1081 cattccctca cacgatacgg taagaagaac caagcgtgag atcgctcaca tgcatgatta
     1141 ccatgattgc tctctgcact tggctcttgc tcctcaggat ggtaaggaag ttttgaagaa
     1201 ggggtggggc cagcgtcacc cactggccgg accaggagtc ccaggccctc ctactgagtg
     1261 gaccttcctt tacgcaccaa ggaacgaaga ggaggccaga gttgtcgaga tgatagtcga
     1321 agctagtatt ggctacatga ctaacgatcc tgctggtaag attgtcgaaa atgccaaagg
     1381 ttgagaccac gaagtggctc ttcagtggac gaaagggcct cgtgatacgc ctatttttat
     1441 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1501 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1561 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg cgctcacgca
     1621 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     1681 cttgttatga ctgttttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     1741 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     1801 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     1861 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     1921 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagcgata ttgatttgct
     1981 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     2041 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     2101 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     2161 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     2221 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     2281 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     2341 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     2401 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     2461 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     2521 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     2581 ttacgtaaaa ggcgagatca ccaaggtagt cggcaaataa ctgtcagacc aagtttactc
     2641 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2701 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2761 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2821 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2881 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2941 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3001 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
//
