LOCUS       sequence-437128-        3963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DVK_AF CIDAR vector encoding mutant of Neonothopanus nambi 
            luciferase nnLuz_v4 under control of CMV promoter, for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3963)
  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 3963)
  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: pNK5664
FEATURES             Location/Qualifiers
     source          1..3963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     464..484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1472..1593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1509..1528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1563..1582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1745..1765
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1766..1823
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1847..1864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2018..2606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2098..2117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2839..3654)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2839..3654)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3562..3581
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3646..3665
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3859..3877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3881..3924)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3927..3948
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggaggcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      121 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      541 cgctaccggt cgccacaatg aggtcaacta tttccttgtc ctcccttctt gaaagattgt
      601 ctaaattgag tagtaggagt atcgctataa cttgtggtgt cgtacttgcc tctgcaatcg
      661 cattccccat aatcaggaga gactaccaga ccttccttga ggtgggtcca agttacgctc
      721 cccagaattt caggggatac ataactgtat gcgtactgtc acttttcaga caagaacaaa
      781 agggtctggc catctacgac agactgcctg aaaaaagaag atggttggct gatttgccct
      841 ttagagaagg accaagacca tccattactt ctcatatcat tcaaaggcaa aggactcaat
      901 tggtagacca ggaatttgca actagggagc tgattgataa ggtgataccc agagtacaag
      961 ctagacatac agacaagact ttcctttcta catctaaatt cgagttccat gctaaggcca
     1021 tcttcctgct tcctagtatt ccaattaacg accccttgaa cattccatcc catgatactg
     1081 tcaggaggac aaagagagaa atcgcccata tgcacgacta ccatgattgt tctttgcacc
     1141 ttgcactggc accacaggac ggcaaggagg tcctgaaaaa gggctggggt caaagacacc
     1201 ccctggcagg tcctggtgta cccggaccac ctactgaatg gacattcttg tatgctccaa
     1261 ggaacgagga ggaggcaagg gtggtggaga tgatcgtaga agcaagtatt ggatatatga
     1321 cgaacgatcc tgctggtaaa attgtggaaa atgctaaagg ttgatgatca taatcagcca
     1381 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1441 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     1501 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1561 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     1621 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1681 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gtcgctatgt
     1741 cttctactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc
     1801 ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact
     1861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc cgcccccctg
     2041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2761 aaacttggtc tgacagctcg agtcccgtca agtcagcgta atgctctgcc agtgttacaa
     2821 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2881 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2941 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3001 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3061 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     3121 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3181 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3241 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3301 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3361 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3421 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3481 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3541 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3601 gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc
     3661 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3721 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     3781 ttttgctgag ttgaaggatc agctcgagtg ccacttgacg tctaagaaac cattattatc
     3841 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc
     3901 ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctagagac tagtggaaga
     3961 cat
//
