LOCUS       sequence-438201-        9489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level M for expression PpASCL in plants 
            (N.benthamiana, BY2 cells).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9489)
  AUTHORS   Palkina KA, Karataeva TA, Perfilov MM, Fakhranurova LI, Markina NM, 
            Somermeyer LG, Garcia-Perez E, Vazquez-Vilar M, Rodriguez-Rodriguez 
            M, Vazquez-Vilriales V, Shakhova ES, Mitiouchkina T, Belozerova OA, 
            Kovalchuk SI, Alekberova A, Malyshevskaia AK, Bugaeva EN, Guglya EB,
            Balakireva A, Sytov N, Bezlikhotnova A, Boldyreva DI, Babenko VV, 
            Kondrashov FA, Choob VV, Orzaez D, Yampolsky IV, Mishin AS, 
            Sarkisyan KS
  TITLE     A hybrid pathway for self-sustained luminescence.
  JOURNAL   Sci Adv. 2024 Mar 8;10(10):eadk1992. doi: 10.1126/sciadv.adk1992. 
            Epub 2024 Mar 8.
  PUBMED    38457503
REFERENCE   2  (bases 1 to 9489)
  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"; journalName: "Sci Adv."; 
            date: "2024-03-8"; volume: "10(10):eadk1992. doi"; pages: " 
            10.1126/sciadv.adk1992. Epub 2024 Mar 8"
COMMENT     Sequence Label: pNK7191
FEATURES             Location/Qualifiers
     source          1..9489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     481..500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             916..2064
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    2210..2234
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2409..2754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2697..2717
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      4126..4833
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        4956..5135
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5171..5226
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     intron          5567..5756
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     terminator      6451..7158
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    7232..7256
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      7344..8054
                     /label=oriV
                     /note="incP origin of replication"
     CDS             8349..9140
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(9481..9489,1..580)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
       61 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      601 tttacggttc ctggcagatc ctagatgtgg cgcaacgatg ccggcgacaa gcaggagcgc
      661 accgacttct tccgcatcaa gtgttttggc tctcaggccg aggcccacgg caagtatttg
      721 ggcaaggggt cgctggtatt cgtgcagggc aagattcgga ataccaagta cgagaaggac
      781 ggccagacgg tctacgggac cgacttcatt gccgataagg tggattatct ggacaccaag
      841 gcaccaggcg ggtcaaatca ggaataaggg cacattgccc cggcgtgagt cggggcaatc
      901 ccgcaaggag ggtgaatgaa tcggacgttt gaccggaagg catacaggca agaactgatc
      961 gacgcggggt tttccgccga ggatgccgaa accatcgcaa gccgcaccgt catgcgtgcg
     1021 ccccgcgaaa ccttccagtc cgtcggctcg atggtccagc aagctacggc caagatcgag
     1081 cgcgacagcg tgcaactggc tccccctgcc ctgcccgcgc catcggccgc cgtggagcgt
     1141 tcgcgtcgtc ttgaacagga ggcggcaggt ttggcgaagt cgatgaccat cgacacgcga
     1201 ggaactatga cgaccaagaa gcgaaaaacc gccggcgagg acctggcaaa acaggtcagc
     1261 gaggccaagc aggccgcgtt gctgaaacac acgaagcagc agatcaagga aatgcagctt
     1321 tccttgttcg atattgcgcc gtggccggac acgatgcgag cgatgccaaa cgacacggcc
     1381 cgctctgccc tgttcaccac gcgcaacaag aaaatcccgc gcgaggcgct gcaaaacaag
     1441 gtcattttcc acgtcaacaa ggacgtgaag atcacctaca ccggcgtcga gctgcgggcc
     1501 gacgatgacg aactggtgtg gcagcaggtg ttggagtacg cgaagcgcac ccctatcggc
     1561 gagccgatca ccttcacgtt ctacgagctt tgccaggacc tgggctggtc gatcaatggc
     1621 cggtattaca cgaaggccga ggaatgcctg tcgcgcctac aggcgacggc gatgggcttc
     1681 acgtccgacc gcgttgggca cctggaatcg gtgtcgctgc tgcaccgctt ccgcgtcctg
     1741 gaccgtggca agaaaacgtc ccgttgccag gtcctgatcg acgaggaaat cgtcgtgctg
     1801 tttgctggcg accactacac gaaattcata tgggagaagt accgcaagct gtcgccgacg
     1861 gcccgacgga tgttcgacta tttcagctcg caccgggagc cgtacccgct caagctggaa
     1921 accttccgcc tcatgtgcgg atcggattcc acccgcgtga agaagtggcg cgagcaggtc
     1981 ggcgaagcct gcgaagagtt gcgaggcagc ggcctggtgg aacacgcctg ggtcaatgat
     2041 gacctggtgc attgcaaacg ctagggcctt gtggggtcag ttccggctgg gggttcagca
     2101 gcccctgctc ggatctgttg gaccggacag tagtcatggt tgatgggctg cctgtatcga
     2161 gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat
     2221 attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa
     2281 tgtactgggg ttgaacactc tgtgggtctc agcaagaatt caagcttgga ggtcaacatg
     2341 gtggagcacg acactctggt ctactccaaa aatgtcaaag atacagtctc agaagatcaa
     2401 agggctattg agacttttca acaaaggata atttcgggaa acctcctcgg attccattgc
     2461 ccagctatct gtcacttcat cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc
     2521 catcattgcg ataaaggaaa ggctatcatt caagatctct ctgccgacag tggtcccaaa
     2581 gatggacccc cacccacgag gagcatcgtg gaaaaagaag aggttccaac cacgtctaca
     2641 aagcaagtgg attgatgtga catctccact gacgtaaggg atgacgcaca atcccactat
     2701 ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag gacacgctcg
     2761 agtataagag ctcattttta caacaattac caacaacaac aaacaacaaa caacattaca
     2821 attacattta caattatcga tacaatggct agtcgtcgtg tcgaggcagc attcgatgga
     2881 caagcagtag aactgggtgc tactattcct gccgccaatg gcaacggaac tcatcagtca
     2941 attaaggttc ctggacatcg tcaagtgact ccaggcaaga ctacgattat ggctattggc
     3001 cgtgctgtgc ccgctaacac gacgttcaat gatggtttgg cagaccatta catccaggag
     3061 ttcaatctgc aagatccagt ccttcaggct aaactgcgtc gtttgtgcga aaccacaacc
     3121 gtgaaaacgc gttatctggt cgtcaataag gagattctgg atgagcatcc tgagttcctg
     3181 gttgatggag ctgcaaccgt atctcagcgt ctggctatta caggcgaagc tgttacccaa
     3241 cttggacatg aggcagcaac ggctgccatc aaagagtggg gccgtccagc ctccgaaatt
     3301 acccatcttg tttatgtttc ctcatccgaa atccgtctgc caggtggcga cttgtacctg
     3361 gcacaattgc ttggcttgcg ttcagacgtg aaccgtgtaa tgttgtacat gctgggttgc
     3421 tatggtggtg catctggaat ccgtgttgcc aaagacttgg ctgagaataa cccaggctca
     3481 cgtgtacttc tgatcacgtc agaatgcact ttgattggat ataagtcact ttcccctgac
     3541 cgtccttacg acctggttgg agcagctctg ttcggtgacg gtgctgccgc tatgattatg
     3601 ggaaaagatc caatccctgt tttggagcgt gctttctttg agcttgactg ggccggtcag
     3661 tcttttattc ctggaactaa caaaaccatc gatggtcgtt tgtctgaaga aggcatttcc
     3721 ttcaagctgg gtcgtgaact gcctaagttg attgaatcca atattcaggg tttctgcgat
     3781 cctatcttga aacgtgctgg tggtctgaag tataacgata ttttctgggc agttcaccca
     3841 ggcggaccag ctatcttgaa tgctgtccag aaacaactgg atttggcacc cgagaaactt
     3901 cagaccgcac gtcaggttct gcgtgactac ggtaacattt ctagttccac ctgcatttac
     3961 gtactggact atatgcgtca tcaatccttg aaactgaagg aagcaaatga caacgtaaat
     4021 acggaaccag agtggggtct gctgcttgca tttggccccg gcgtcactat cgagggtgct
     4081 ttgctgcgta acctgtgctc aggtagttaa ctagcttaat aggtcctgct ttaatgagat
     4141 atgcgagaag cctatgatcg catgatattt gctttcaatt ctgttgtgca cgttgtaaaa
     4201 aacctgagca tgtgtagctc agatccttac cgccggtttc ggttcattct aatgaatata
     4261 tcacccgtta ctatcgtatt tttatgaata atattctccg ttcaatttac tgattgtacc
     4321 ctactactta tatgtacaat attaaaatga aaacaatata ttgtgctgaa taggtttata
     4381 gcgacatcta tgatagagcg ccacaataac aaacaattgc gttttattat tacaaatcca
     4441 attttaaaaa aagcggcaga accggtcaaa cctaaaagac tgattacata aatcttattc
     4501 aaatttcaaa agtgccccag gggctagtat ctacgacaca ccgagcggcg aactaataac
     4561 gctcactgaa gggaactccg gttccccgcc ggcgcgcatg ggtgagattc cttgaagttg
     4621 agtattggcc gtccgctcta ccgaaagtta cgggcaccat tcaacccggt ccagcacggc
     4681 ggccgggtaa ccgacttgct gccccgagaa ttatgcagca tttttttggt gtatgtgggc
     4741 cccaaatgaa gtgcaggtca aaccttgaca gtgacgacaa atcgttgggc gggtccaggg
     4801 cgaattttgc gacaacatgt cgaggctcag caggaccgct actagcaaca tacgctggat
     4861 tacgaattcc catggggaga gcggagaatt aagggagtca cgttatgacc cccgccgatg
     4921 acgcgggaca agccgtttta cgtttggaac tgacagaacc gcaacgttga aggagccact
     4981 cagccgcggg tttctggagt ttaatgagct aagcacatac gtcagaaacc attattgcgc
     5041 gttcaaaagt cgcctaaggt cactatcagc tagcaaatat ttcttgtcaa aaatgctcca
     5101 ctgacgttcc ataaattccc ctcggtatcc aattagagtc tcatattcac tctcatactg
     5161 tatttttaca acaattacca acaacaacaa acaacaaaca acattacaat tactatttac
     5221 aattacaatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     5281 gttcgacagt gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     5341 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     5401 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     5461 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     5521 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcaggtaa atttctagtt
     5581 tttctccttc attttcttgg ttaggaccct tttctctttt tatttttttg agctttgatc
     5641 tttctttaaa ctgatctatt ttttaattga ttggttatgg tgtaaatatt acatagcttt
     5701 aactgataat ctgattactt tatttcgtgt gtctatgatg atgatgataa ctgcagccgg
     5761 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     5821 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     5881 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     5941 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     6001 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     6061 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     6121 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     6181 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     6241 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     6301 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     6361 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     6421 ctcgtccgag ggcaaaggaa taggcttgtc ctgctttaat gagatatgcg agaagcctat
     6481 gatcgcatga tatttgcttt caattctgtt gtgcacgttg taaaaaacct gagcatgtgt
     6541 agctcagatc cttaccgccg gtttcggttc attctaatga atatatcacc cgttactatc
     6601 gtatttttat gaataatatt ctccgttcaa tttactgatt gtaccctact acttatatgt
     6661 acaatattaa aatgaaaaca atatattgtg ctgaataggt ttatagcgac atctatgata
     6721 gagcgccaca ataacaaaca attgcgtttt attattacaa atccaatttt aaaaaaagcg
     6781 gcagaaccgg tcaaacctaa aagactgatt acataaatct tattcaaatt tcaaaagtgc
     6841 cccaggggct agtatctacg acacaccgag cggcgaacta ataacgctca ctgaagggaa
     6901 ctccggttcc ccgccggcgc gcatgggtga gattccttga agttgagtat tggccgtccg
     6961 ctctaccgaa agttacgggc accattcaac ccggtccagc acggcggccg ggtaaccgac
     7021 ttgctgcccc gagaattatg cagcattttt ttggtgtatg tgggccccaa atgaagtgca
     7081 ggtcaaacct tgacagtgac gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa
     7141 catgtcgagg ctcagcagga ccgctcagat gagaccgagg atgcacatgt gaccgaggga
     7201 cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa
     7261 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
     7321 ttcgtccatt tgtatgtgcc agccgccttt gcgacgctca ccgggctggt tgccctcgcc
     7381 gctgggctgg cggccgtcta tggccctgca aacgcgccag aaacgccgtc gaagccgtgt
     7441 gcgagacacc gcggccgccg gcgttgtgga tacctcgcgg aaaacttggc cctcactgac
     7501 agatgagggg cggacgttga cacttgaggg gccgactcac ccggcgcggc gttgacagat
     7561 gaggggcagg ctcgatttcg gccggcgacg tggagctggc cagcctcgca aatcggcgaa
     7621 aacgcctgat tttacgcgag tttcccacag atgatgtgga caagcctggg gataagtgcc
     7681 ctgcggtatt gacacttgag gggcgcgact actgacagat gaggggcgcg atccttgaca
     7741 cttgaggggc agagtgctga cagatgaggg gcgcacctat tgacatttga ggggctgtcc
     7801 acaggcagaa aatccagcat ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct
     7861 gtcttttaac ctgcttttaa accaatattt ataaaccttg tttttaacca gggctgcgcc
     7921 ctgtgcgcgt gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac cctcccggcc
     7981 cgctaacgcg ggcctcccat ccccccaggg gctgcgcccc tcggccgcga acggcctcac
     8041 cccaaaaatg gcagcgctgg ccaattcccg aggcacgaac ccagtggaca taagcctgtt
     8101 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     8161 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     8221 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     8281 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     8341 taaacatcat gggggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     8401 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     8461 tggatggcgg cctgaagcca cacagcgata ttgatttgct ggttacggtg accgtaaggc
     8521 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     8581 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     8641 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     8701 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     8761 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     8821 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     8881 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     8941 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     9001 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     9061 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ttacgtgaaa ggcgagatca
     9121 ccaaggtagt cggcaaataa tgtctagcta gaaattcgtt caagccgacg ccgcttcgcg
     9181 gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag ccaaactatc
     9241 aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac aaattgggag
     9301 atatatcatg ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     9421 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     9481 ttgagatcc
//
