LOCUS       sequence-437134-        8209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DVK_AF CIDAR vector encoding hispidin-synthase from Mycena 
            citricolor under control of CMV promoter, for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8209)
  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 8209)
  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: pNK3203
FEATURES             Location/Qualifiers
     source          1..8209
                     /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    5718..5839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5755..5774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5809..5828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    5991..6011
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      6012..6069
                     /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(6093..6110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6264..6852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6344..6363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7085..7900)
                     /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(7085..7900)
                     /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     7808..7827
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7892..7911
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     8105..8123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(8127..8170)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    8173..8194
                     /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 tcagcctctt catcttattc tgaattggag tctccaactt
      601 ctcttcttga cgtgttcgtg cacgccgctc gggaccctca caccgcatca agaagggtgt
      661 tggagtgcgg atctgacaca tggacatacg ccgccttgga tgccgtgtct gatggtatag
      721 ccagggagtt ggcccctttc ggtttggccc caaaggtcgc tgttgtctct gagaaccatc
      781 catttgtgtt cgctttgttg ttcgcagtct ggaagttggg tggaactttc atcccaatcg
      841 acgcacatgt cccttttgcc atgttgacag gtatggtgaa catcgtgaag ccaacatgtc
      901 tttaccttcc tgcctcagcc acatcaaaca tatctcttgt gaaggccttc gacataagaa
      961 ccgtcgtgtt tggacacaag gagaattcaa tgcaagcttt gttcgacaag tactcattgc
     1021 acgcagcccc attgcacgct gcaccattgc attacgcccc tccatcagcc gaccatgcct
     1081 gtctttatct tttcacttct tcagcctctt ctaccaaaaa cttgaaggcc gtccctctta
     1141 cccatacact tgtcttgaga ggttgtcaat ctaaacttgc ttggtggagg agggtccaac
     1201 caggaaagaa tcttgacgct atcagaatcc ttggatgggc accatggtcc cacgttcttg
     1261 cctatatgca ggatatcgga acagctacac ttcttaacgc cggttgctac gtctttgcaa
     1321 ccatcccttc ttcataccct cagcttccta ccactacccc agtcgatttg accacttcac
     1381 ttatcgacgc tttggtcacc agaagagtcg cagcattcgc ctgtgtccct ttcgtccttg
     1441 ccaaccttaa agccgcttgc cagtcaaacc acccagccag gtcacagttg cttgaggccc
     1501 ttcaaaagac catcatgttg gagtgcggtg gtgccgtgtt ggacgacgct actgtggact
     1561 gggccgagag gaacggtatc aggatcttca ccggaatcgg tatgaccgag actggtggag
     1621 ccgttttcgt cggtttggcc tctgaatcta gaagaggatt tttgcctgaa ggtttgcttg
     1681 gtgatgcatc attctctatc tcatctgaca ctgacgctct tgacgaaggt gaacttgtcg
     1741 tgaaatcaaa attgatcgca ccaggatatg tcggatacga cgacggagcc cactctgtgg
     1801 actgcgacgg atgggtcaca ttcaggacag gagataggta taggcagacc cagccagacg
     1861 gaaggtttac ctggttggga aggatcacag acttcattca gatggtgtct ggtgagtact
     1921 tggatccaag gccattggag gagtctctta gagcctctcc attgatcgcc aacgcctgca
     1981 tcgtgggtga cgccttccta tcttctgcct caacttcaat cttggccatc atcgagttgg
     2041 caacccctga tcttgcacac accgcctcaa tcagggcaca gcttgcaaga gtccttgccc
     2101 ctcttaacag ggatttgcct ccaccactta ggatcgcagc atcttcaatc cttgttttgg
     2161 acggaatgag aaagatccct aagacaaaga agggtgacat cttcagaaaa aagcttgagg
     2221 acacattcgg tgctgagttt gagcagatgt tgagaaccga gaaagtcgga ttgggtgact
     2281 tggctgatgt ggatgccggt atcactagaa tcatcggaaa cttgttggga atctcagacg
     2341 acgagcttct ttctaccatg tcttttgctg agttgggtat gacctctctt ttggccgtca
     2401 agatcgcatc agaattgaac aagtttttgg acggtagggc tgtgttgcca acaaacatct
     2461 gttacataca ctttgacgtg ccttcattga cttcatcagt cagagaaagg ctttcttctg
     2521 caccatcatc tatcacatct gccgcctctg agcctgcagc ctcttcacca ggaaccccta
     2581 gggccgacga aatcgtgatc gtgggaaagg ccttcagact tcctgacggt gtgaacgacg
     2641 acgccgcttt gtgggacgtc cttaccggtg agtctgcatc tatcatcaag gacatcccag
     2701 ccgacagatg ggaccacgcc tctttctacc caaaggacat ccactttggg agagctggtc
     2761 ttgtggatgt cgctagattc gactacggtt ttttcggaat gaccgcttct gaggcatatt
     2821 ctctttcacc aaccatgagg ttggctcttg aagtcgctta tgaggccttg gaggacgcaa
     2881 acattccttt cagagccgtc aagggttcaa ggatgggtgt gttcgtcgct gtgaaagacg
     2941 atggattcga aaccttgttg catgcaggac agggttatga cgcttacaca agattctatg
     3001 gaactggaag ggctccttca accgcctctg gtagaatcaa ctatttgctt gatttgcacg
     3061 gaccttcaat caccgtcgac acagcctgct caggaggaat cgtgtgcatc gaccaggctg
     3121 tcacctatct tcaatcagga gctgcagaga ctgccatcgt gtgttcatct aacacacact
     3181 gctggccagg atctttcatg ttcttgaccg ctcaaggaat ggcctcacct aacggtagat
     3241 gcgcatcttt cacatcagac gccgacggat acgctccttc tgagggagct gtgggtttcg
     3301 tgcttaagac caggtctgcc gctgtgaggg acggagatag aatacttgct actatcaggg
     3361 ccaccgagat cggacacaat ggaagatctc agggacttgc tgctcctaat gtcagatctc
     3421 aggcagctgc tcatagagcc gtgttgcgta gagcaaggtt ggacccatct gagattgact
     3481 tcatcgaagc acatggtact ggaaccacat tgggagatct ttgcgaagtg cagggtatca
     3541 acgactcttt tgtctctcct aagaaaagag ccaaccctct tgtagtctct gcatctaagt
     3601 caaccatcgg tcacaccgag ccatctgctg gtttggtcgg aatcctttca gctcttatgt
     3661 cattcgagaa gaggatcgtg cctagattgg catatcttac tgagtctaat gtgaacccag
     3721 cattggacgc atcagtcgtt ccattgcact ttcctacaaa gcacatcgag cttagagctg
     3781 atgtgcctta caaggccgtg gtgatgtctt acggattcgc aggtaccctt gccgacatcg
     3841 tccttgagtc agaggttcca caaccaactc cagccgtcgc tcaggacaca gccggtcaac
     3901 agccaatgct tttcgtgttg tcagccaaaa ccccaagggc acttgcagcc tacatcgagc
     3961 tttaccttgg tttccttagg cacgccgacc ctggactttt cgcaaggatc tgctacaccg
     4021 catgtgtcgc aagggagcac tacaagcaca gggtcgcctg cgttgcaaca gacttggtcg
     4081 accttatcgc tcagcttgag actaggcttg tgcagactgc ctatgcagga ggaggaggtg
     4141 ccagggccgc taggaccggt cctcttgtgt ttgctttttc tggacagggt actcagttcc
     4201 ctgctatggc agcaccactt gctaggagat acgcaagatt cggagagatc gtgggtggtt
     4261 gtgctaggat ggcaagagag ctttcaggtt tcccagtgga tggtattctt cttggtgacg
     4321 atgtgacacc tgtgaaggac aactctgctg cagctgaggt gcactctgag gtggaccaga
     4381 tctgcatatt tgtgtaccag tatgccatgt gtagatggtt gggtgagctt ggtgtggagc
     4441 caaaggctgc cataggtcac tctttgggag agatcacagc agctgtcatc gcaggagcac
     4501 ttcctttcga agccgccttg gatcttgtcg tcactagagc caggttgctt aagccatgcg
     4561 ccgagcaacc ttctggaatg gcagccttgg cctgcacccc agacgtcgca tcaaagttga
     4621 cacttggtgc ttcagtgtct gtgtcagtct acaacggacc tcagtctatc tgcctttctg
     4681 gtgcttctgc cgagcttgac gatgccgtga gggccgctaa ggccaggaac atcaaggcaa
     4741 ctagacttca ggtcgaccag ggtttccact caccttgcgt cgacgccgca gtgccaggtt
     4801 tgcaggcttg gtgcgcagct cacagggcat cagctgctcc attgaagatg cctctttact
     4861 ctaccgtgag aggggacgtc gttccaaagg gagccgccct tgatccagag cactgggtgg
     4921 cccacgctag gaaccctgtg ttgttcgccc agacagccca ggctcttaaa gagtcacttc
     4981 cacatggaat cactttggac gtcggtcctc aggccgtggc ttggtctttg cttcttttga
     5041 acggtctttc agctaccaga acagtcgctg ccggagccaa gaagggagca gaccaggaga
     5101 gggccctttt gggtgctctt ggagccttgt tcgagcagca caaggtcact cctgactttg
     5161 gtagacttta cgctcctttg gaaaagacaa ggatcccaac ctacccattt gagagagcta
     5221 gatgctaccc aaccttcata ccatcaaggt tcgcccacgg tgctgctgct gctgccaaga
     5281 cagatggtga ggtcctttct gccgaagaag aaaatgtggc acctgtggga ttgttgacaa
     5341 aggaggattt gagggcagct ttggtcgctt gtttgagggc cacattggag cttaggcctg
     5401 acgaggagct tgacgaagca gagcctttga ccgtgcacgg agtggactct atcggatttg
     5461 caaagcttag gaagcacgtc gaggacaggt ggggtcttga catcccagtc gtgtactggt
     5521 cagacgcctt caccgtcgga gagatgttgg gtaacttggt cggacagtat gacgtggtgt
     5581 ctactgctgc caggtagtta actagcttaa tgatcataat cagccatacc acatttgtag
     5641 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5701 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5821 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     5881 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5941 tataaatcaa aagaatagac cgagataggg ttgagtgtcg ctatgtcttc tactagtagc
     6001 ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa
     6061 ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc
     6121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6181 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6241 aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa
     6301 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6421 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6601 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6661 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6721 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6781 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6841 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6901 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6961 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7021 agctcgagtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     7081 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     7141 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     7201 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     7261 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     7321 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     7381 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     7441 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     7501 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     7561 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     7621 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     7681 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     7741 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     7801 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     7861 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     7921 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     7981 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     8041 aggatcagct cgagtgccac ttgacgtcta agaaaccatt attatcatga cattaaccta
     8101 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
     8161 ggatgatttc tggaattcgc ggccgcttct agagactagt ggaagacat
//
