LOCUS       sequence-437130-        4428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DVK_AF CIDAR vector encoding mutant of Neonothopanus nambi 
            hispidin-3-hydroxylase nnH3H_v2 under control of CMV promoter, for 
            mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4428)
  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 4428)
  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: pNK291
FEATURES             Location/Qualifiers
     source          1..4428
                     /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    1937..2058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1974..1993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2028..2047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2210..2230
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2231..2288
                     /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(2312..2329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2483..3071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2563..2582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3304..4119)
                     /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(3304..4119)
                     /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     4027..4046
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4111..4130
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4324..4342
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4346..4389)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4392..4413
                     /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 gcttctttcg agaacagcct ttccgttctt atagtgggtg
      601 cgggcctggg tggattggcg gctgcgatag cactgcgaag gcagggccac gttgtaaaaa
      661 tatatgaatc atcctctttc aaggctgagt tgggtgccgg gcttgcggtg cctcccaaca
      721 ccttgaggtc attgcaacag cttgggtgca atacagaaaa cctgaatggc gtcgataatt
      781 tgtgtttcac ggctatgggc tacgatggaa gtgttgggat gatgaacaat atgaccgact
      841 acagggaggc ctatggcacg tcatggataa tggtacatag ggttgatttg cataacgaac
      901 ttatgagggt ggcccttgat cctgggggcc tcggtccccc cgcaaccctg catctcaacc
      961 atagggttac tttttgcgat gtcgatgcct gtaccgtaac atttaccaac ggaaccactc
     1021 agtcagccga tttgattgtt ggtgcagatg gaataagatc cacgattcgg cgcttcgtgc
     1081 tcgaagagga cgtcacaatc ccagcttctg gcatagtagg ttttagatgg cttgtgcagg
     1141 ctgatgcact cgatccgtat ccagaactcg actggatagt aaagaaacct ccactcggag
     1201 caagactcat ctctacaccg cagaatccac agagtggggt ggggttggcc gaccggcgga
     1261 caattatcat ctatgcatgc cgagggggta cgatggtcaa tgttctcgct gtgcatgacg
     1321 atgaaagaga tcagaatacg gcagactggt cagtgcccgc gagtaaggat gacctcttta
     1381 gggtgtttca tgattatcac ccacggttca gacgcctgct ggaattggcc caggacatca
     1441 atctctggca aatgcgcgtc gtcccggtcc tcaagaaatg ggtgaataag cgcgtgtgcc
     1501 tgctcggcga tgcagcgcat gccatgctcc ccacgctggg ccaaggtttc gggatgggcc
     1561 tcgaggatgc cgtggcgctt ggcacgcttt tgccaaaggg gaccactgcc agtcagatcg
     1621 aaacaagatt ggctgtttat gaacagctga ggaaggaccg agcggagttt gtggcagcag
     1681 agagttacga agaacagtat gtcccggaga aacgcggact ttacctgaga agcaaagaac
     1741 ttagagatcg cgtcatggga tacgatatca aggtggagtc agagaaagtg ttggaaacgc
     1801 ttttgcgaag ctctaatagc gcaggttgat gatcataatc agccatacca catttgtaga
     1861 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1921 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2041 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2101 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2161 ataaatcaaa agaatagacc gagatagggt tgagtgtcgc tatgtcttct actagtagcg
     2221 gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac
     2281 cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg
     2341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2461 aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa
     2521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2641 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2821 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2881 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3001 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3181 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3241 gctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     3301 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     3361 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3421 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     3481 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3541 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     3601 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3661 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     3721 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3781 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3841 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3901 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     3961 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     4021 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     4081 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     4141 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     4201 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     4261 ggatcagctc gagtgccact tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4321 aaaaataggc gtatcacgag gcagaatttc agataaaaaa aatccttagc tttcgctaag
     4381 gatgatttct ggaattcgcg gccgcttcta gagactagtg gaagacat
//
