LOCUS       sequence-437095-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level 0 promoterless vector encoding 
            hispidin-synthase from Mycena citricolor codon-optimised for 
            expression in Nicotiana benthamiana, Pichia pastoris, Homo sapiens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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: pNK2657
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      287..875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     776..795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1029..1046
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6197..6215)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6283..6387
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6609..7284,1..116)
                     /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 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
       61 tggaagaatt tgtccattac gtaaaaggcg agatcaccaa ggtagtcggc aaataactgt
      121 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      901 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      961 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1021 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     1081 tccccgcgcg ttggccgatt cattaatcac tctgtggtct caaatgtcag cctcttcatc
     1141 ttattctgaa ttggagtctc caacttctct tcttgacgtg ttcgtgcacg ccgctcggga
     1201 ccctcacacc gcatcaagaa gggtgttgga gtgcggatct gacacatgga catacgccgc
     1261 cttggatgcc gtgtctgatg gtatagccag ggagttggcc cctttcggtt tggccccaaa
     1321 ggtcgctgtt gtctctgaga accatccatt tgtgttcgct ttgttgttcg cagtctggaa
     1381 gttgggtgga actttcatcc caatcgacgc acatgtccct tttgccatgt tgacaggtat
     1441 ggtgaacatc gtgaagccaa catgtcttta ccttcctgcc tcagccacat caaacatatc
     1501 tcttgtgaag gccttcgaca taagaaccgt cgtgtttgga cacaaggaga attcaatgca
     1561 agctttgttc gacaagtact cattgcacgc agccccattg cacgctgcac cattgcatta
     1621 cgcccctcca tcagccgacc atgcctgtct ttatcttttc acttcttcag cctcttctac
     1681 caaaaacttg aaggccgtcc ctcttaccca tacacttgtc ttgagaggtt gtcaatctaa
     1741 acttgcttgg tggaggaggg tccaaccagg aaagaatctt gacgctatca gaatccttgg
     1801 atgggcacca tggtcccacg ttcttgccta tatgcaggat atcggaacag ctacacttct
     1861 taacgccggt tgctacgtct ttgcaaccat cccttcttca taccctcagc ttcctaccac
     1921 taccccagtc gatttgacca cttcacttat cgacgctttg gtcaccagaa gagtcgcagc
     1981 attcgcctgt gtccctttcg tccttgccaa ccttaaagcc gcttgccagt caaaccaccc
     2041 agccaggtca cagttgcttg aggcccttca aaagaccatc atgttggagt gcggtggtgc
     2101 cgtgttggac gacgctactg tggactgggc cgagaggaac ggtatcagga tcttcaccgg
     2161 aatcggtatg accgagactg gtggagccgt tttcgtcggt ttggcctctg aatctagaag
     2221 aggatttttg cctgaaggtt tgcttggtga tgcatcattc tctatctcat ctgacactga
     2281 cgctcttgac gaaggtgaac ttgtcgtgaa atcaaaattg atcgcaccag gatatgtcgg
     2341 atacgacgac ggagcccact ctgtggactg cgacggatgg gtcacattca ggacaggaga
     2401 taggtatagg cagacccagc cagacggaag gtttacctgg ttgggaagga tcacagactt
     2461 cattcagatg gtgtctggtg agtacttgga tccaaggcca ttggaggagt ctcttagagc
     2521 ctctccattg atcgccaacg cctgcatcgt gggtgacgcc ttcctatctt ctgcctcaac
     2581 ttcaatcttg gccatcatcg agttggcaac ccctgatctt gcacacaccg cctcaatcag
     2641 ggcacagctt gcaagagtcc ttgcccctct taacagggat ttgcctccac cacttaggat
     2701 cgcagcatct tcaatccttg ttttggacgg aatgagaaag atccctaaga caaagaaggg
     2761 tgacatcttc agaaaaaagc ttgaggacac attcggtgct gagtttgagc agatgttgag
     2821 aaccgagaaa gtcggattgg gtgacttggc tgatgtggat gccggtatca ctagaatcat
     2881 cggaaacttg ttgggaatct cagacgacga gcttctttct accatgtctt ttgctgagtt
     2941 gggtatgacc tctcttttgg ccgtcaagat cgcatcagaa ttgaacaagt ttttggacgg
     3001 tagggctgtg ttgccaacaa acatctgtta catacacttt gacgtgcctt cattgacttc
     3061 atcagtcaga gaaaggcttt cttctgcacc atcatctatc acatctgccg cctctgagcc
     3121 tgcagcctct tcaccaggaa cccctagggc cgacgaaatc gtgatcgtgg gaaaggcctt
     3181 cagacttcct gacggtgtga acgacgacgc cgctttgtgg gacgtcctta ccggtgagtc
     3241 tgcatctatc atcaaggaca tcccagccga cagatgggac cacgcctctt tctacccaaa
     3301 ggacatccac tttgggagag ctggtcttgt ggatgtcgct agattcgact acggtttttt
     3361 cggaatgacc gcttctgagg catattctct ttcaccaacc atgaggttgg ctcttgaagt
     3421 cgcttatgag gccttggagg acgcaaacat tcctttcaga gccgtcaagg gttcaaggat
     3481 gggtgtgttc gtcgctgtga aagacgatgg attcgaaacc ttgttgcatg caggacaggg
     3541 ttatgacgct tacacaagat tctatggaac tggaagggct ccttcaaccg cctctggtag
     3601 aatcaactat ttgcttgatt tgcacggacc ttcaatcacc gtcgacacag cctgctcagg
     3661 aggaatcgtg tgcatcgacc aggctgtcac ctatcttcaa tcaggagctg cagagactgc
     3721 catcgtgtgt tcatctaaca cacactgctg gccaggatct ttcatgttct tgaccgctca
     3781 aggaatggcc tcacctaacg gtagatgcgc atctttcaca tcagacgccg acggatacgc
     3841 tccttctgag ggagctgtgg gtttcgtgct taagaccagg tctgccgctg tgagggacgg
     3901 agatagaata cttgctacta tcagggccac cgagatcgga cacaatggaa gatctcaggg
     3961 acttgctgct cctaatgtca gatctcaggc agctgctcat agagccgtgt tgcgtagagc
     4021 aaggttggac ccatctgaga ttgacttcat cgaagcacat ggtactggaa ccacattggg
     4081 agatctttgc gaagtgcagg gtatcaacga ctcttttgtc tctcctaaga aaagagccaa
     4141 ccctcttgta gtctctgcat ctaagtcaac catcggtcac accgagccat ctgctggttt
     4201 ggtcggaatc ctttcagctc ttatgtcatt cgagaagagg atcgtgccta gattggcata
     4261 tcttactgag tctaatgtga acccagcatt ggacgcatca gtcgttccat tgcactttcc
     4321 tacaaagcac atcgagctta gagctgatgt gccttacaag gccgtggtga tgtcttacgg
     4381 attcgcaggt acccttgccg acatcgtcct tgagtcagag gttccacaac caactccagc
     4441 cgtcgctcag gacacagccg gtcaacagcc aatgcttttc gtgttgtcag ccaaaacccc
     4501 aagggcactt gcagcctaca tcgagcttta ccttggtttc cttaggcacg ccgaccctgg
     4561 acttttcgca aggatctgct acaccgcatg tgtcgcaagg gagcactaca agcacagggt
     4621 cgcctgcgtt gcaacagact tggtcgacct tatcgctcag cttgagacta ggcttgtgca
     4681 gactgcctat gcaggaggag gaggtgccag ggccgctagg accggtcctc ttgtgtttgc
     4741 tttttctgga cagggtactc agttccctgc tatggcagca ccacttgcta ggagatacgc
     4801 aagattcgga gagatcgtgg gtggttgtgc taggatggca agagagcttt caggtttccc
     4861 agtggatggt attcttcttg gtgacgatgt gacacctgtg aaggacaact ctgctgcagc
     4921 tgaggtgcac tctgaggtgg accagatctg catatttgtg taccagtatg ccatgtgtag
     4981 atggttgggt gagcttggtg tggagccaaa ggctgccata ggtcactctt tgggagagat
     5041 cacagcagct gtcatcgcag gagcacttcc tttcgaagcc gccttggatc ttgtcgtcac
     5101 tagagccagg ttgcttaagc catgcgccga gcaaccttct ggaatggcag ccttggcctg
     5161 caccccagac gtcgcatcaa agttgacact tggtgcttca gtgtctgtgt cagtctacaa
     5221 cggacctcag tctatctgcc tttctggtgc ttctgccgag cttgacgatg ccgtgagggc
     5281 cgctaaggcc aggaacatca aggcaactag acttcaggtc gaccagggtt tccactcacc
     5341 ttgcgtcgac gccgcagtgc caggtttgca ggcttggtgc gcagctcaca gggcatcagc
     5401 tgctccattg aagatgcctc tttactctac cgtgagaggg gacgtcgttc caaagggagc
     5461 cgcccttgat ccagagcact gggtggccca cgctaggaac cctgtgttgt tcgcccagac
     5521 agcccaggct cttaaagagt cacttccaca tggaatcact ttggacgtcg gtcctcaggc
     5581 cgtggcttgg tctttgcttc ttttgaacgg tctttcagct accagaacag tcgctgccgg
     5641 agccaagaag ggagcagacc aggagagggc ccttttgggt gctcttggag ccttgttcga
     5701 gcagcacaag gtcactcctg actttggtag actttacgct cctttggaaa agacaaggat
     5761 cccaacctac ccatttgaga gagctagatg ctacccaacc ttcataccat caaggttcgc
     5821 ccacggtgct gctgctgctg ccaagacaga tggtgaggtc ctttctgccg aagaagaaaa
     5881 tgtggcacct gtgggattgt tgacaaagga ggatttgagg gcagctttgg tcgcttgttt
     5941 gagggccaca ttggagctta ggcctgacga ggagcttgac gaagcagagc ctttgaccgt
     6001 gcacggagtg gactctatcg gatttgcaaa gcttaggaag cacgtcgagg acaggtgggg
     6061 tcttgacatc ccagtcgtgt actggtcaga cgccttcacc gtcggagaga tgttgggtaa
     6121 cttggtcgga cagtatgacg tggtgtctac tgctgccagg ttgagaccac gaagtggctc
     6181 ttcagtggac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6241 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6361 cttcaataat attgaaaaag gaagagtatg cgctcacgca actggtccag aaccttgacc
     6421 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     6481 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     6541 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     6601 taaacatcat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     6661 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     6721 tggatggcgg cctgaagcca cacagcgata ttgatttgct ggttacggtg accgtaaggc
     6781 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     6841 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     6901 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     6961 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     7021 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     7081 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     7141 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     7201 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     7261 cccagtatca gcccgtcata cttg
//
