LOCUS       sequence-437298-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level 1 for expression HmS in plants 
            (N.benthamiana, BY2 cells).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  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: pNK7180
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     302..321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(540..644)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      731..1362
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1454..1478
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        1664..2009
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1952..1972
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      3324..4031
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4080..4104
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4260..5408)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(5744..6332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5824..5843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6503..6824,1..470,471..539))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
       61 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
      121 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
      181 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
      241 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
      301 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
      361 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
      421 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
      481 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
      541 ctcttccttt ttcaatatta ttgaagcatt tatcaaggtt attgtctcat gagcggatac
      601 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacgaa ttggccagcg
      661 ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga
      721 ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc
      781 ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa
      841 gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg
      901 gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca
      961 ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag
     1021 tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg
     1081 taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat
     1141 cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg
     1201 tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg
     1261 ccgcggtgtc tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg gccatagacg
     1321 gccgccagcc cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc
     1381 tgactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct
     1441 gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta
     1501 ataacacatt gcggacgttt ttaatgtact ggggtggatg cagtgggccc cactctgtga
     1561 agacaagcaa gaattcaagc ttggaggtca acatggtgga gcacgacact ctggtctact
     1621 ccaaaaatgt caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa
     1681 ggataatttc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa
     1741 ggacagtaga aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta
     1801 tcattcaaga tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
     1861 tcgtggaaaa agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct
     1921 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     1981 aaggaagttc atttcatttg gagaggacac gctcgagtat aagagctcat ttttacaaca
     2041 attaccaaca acaacaaaca acaaacaaca ttacaattac atttacaatt atcgatacaa
     2101 tggctaccaa aagtgtcgcc gttgaagaaa tgtgcaaagc ccaaaaagcc ggaggccctg
     2161 caacaatcct tgcaattggt actgctgtgc ccagtaactg ctactatcag agtgaatacc
     2221 ctgatttcta cttccgtgtc acgaaaagtg atcaccttac ggacctgaaa agtaagttta
     2281 aacgtatgtg cgaacgttca tccatcaaaa agcgttatat gcacctgaca gaggagattc
     2341 ttgaagaaaa tcctaatatg tgcacgtttg cagctccatc tatcgatggc cgtcaggaca
     2401 ttgtagttaa ggagatccct aagttggcta aggaggcagc aagtaaagct atcaaggaat
     2461 ggggccaacc taagtcaaat atcacccatc ttgtcttttg tacaacttca ggtgtggaca
     2521 tgcccggttg tgattatcaa ttgacgcgtc ttttgggact tcgtccatct attaaacgtc
     2581 ttatgatgta tcagcagggc tgccatgctg gtggaaccgg cttgcgtttg gccaaagacc
     2641 ttgccgaaaa taacaaggga gctcgtgtat tggtggtttg ttctgagatg acagtgatta
     2701 actttcgtgg ccccagtgag gcccacatgg attcccttgt cggacaatct cttttcggcg
     2761 acggcgcctc agccgtaatc gtaggttcag atcccgatct gtcaacagag catccacttt
     2821 accagattat gtccgcctcc caaatcattg tagccgactc cgaaggcgct attgatggtc
     2881 acttgcgtca agagggtctt acctttcatc tgcgtaagga tgttccttct cttgtctccg
     2941 ataacatcga gaatactctg gtggaagcat ttactcccat cctgatggac agtatcgact
     3001 caatcattga ttggaactct atcttttgga ttgctcaccc aggtggtcct gccatcctga
     3061 atcaggttca agcaaaagta ggactgaagg aggaaaaact gcgtgtgagt cgtcacattt
     3121 tgtcagaata tggcaatatg tcatccgcat gcgtcttttt tatcatggat gagatgcgta
     3181 aacgttccat ggaagaggga aaaggaacta ccggcgaggg ccttgagtgg ggagtgcttt
     3241 ttggcttcgg tcctggtttt acggtcgaaa ccattgtgtt gcacagtgtg cccatttcag
     3301 gtagttaact agcttaatag gtcctgcttt aatgagatat gcgagaagcc tatgatcgca
     3361 tgatatttgc tttcaattct gttgtgcacg ttgtaaaaaa cctgagcatg tgtagctcag
     3421 atccttaccg ccggtttcgg ttcattctaa tgaatatatc acccgttact atcgtatttt
     3481 tatgaataat attctccgtt caatttactg attgtaccct actacttata tgtacaatat
     3541 taaaatgaaa acaatatatt gtgctgaata ggtttatagc gacatctatg atagagcgcc
     3601 acaataacaa acaattgcgt tttattatta caaatccaat tttaaaaaaa gcggcagaac
     3661 cggtcaaacc taaaagactg attacataaa tcttattcaa atttcaaaag tgccccaggg
     3721 gctagtatct acgacacacc gagcggcgaa ctaataacgc tcactgaagg gaactccggt
     3781 tccccgccgg cgcgcatggg tgagattcct tgaagttgag tattggccgt ccgctctacc
     3841 gaaagttacg ggcaccattc aacccggtcc agcacggcgg ccgggtaacc gacttgctgc
     3901 cccgagaatt atgcagcatt tttttggtgt atgtgggccc caaatgaagt gcaggtcaaa
     3961 ccttgacagt gacgacaaat cgttgggcgg gtccagggcg aattttgcga caacatgtcg
     4021 aggctcagca ggaccgctac tattgtcttc tgcacgaagt ggtttaaact atcagtgttt
     4081 gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata
     4141 tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag
     4201 ggttccccag atcaggcgct ggctgctgaa cccccagccg gaactgaccc cacaaggccc
     4261 tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct
     4321 cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc
     4381 gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg
     4441 aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat
     4501 ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa
     4561 cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc
     4621 aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat
     4681 tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg
     4741 tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caacacctgc
     4801 tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg
     4861 tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg
     4921 cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc
     4981 cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc
     5041 agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt
     5101 cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc
     5161 gcctcctgtt caagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg
     5221 ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac catcgagccg
     5281 acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca
     5341 tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc
     5401 cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat gtgcccttat
     5461 tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc ggcaatgaag
     5521 tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat cttgccctgc
     5581 acgaatacca gcgacccctt gcccaaatac ttgccgtggg cctcggcctg agagccaaaa
     5641 cacttgatgc ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc
     5701 taggatctgc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6181 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg
     6661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggc
//
