LOCUS       sequence-437297-        6881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level 1 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 6881)
  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 6881)
  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: pNK7187
FEATURES             Location/Qualifiers
     source          1..6881
                     /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      3381..4088
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4137..4161
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4317..5465)
                     /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(5801..6389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5881..5900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6560..6881,1..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 agacaagagc gaattccata tgggaggtca 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 tggctagtcg tcgtgtcgag gcagcattcg atggacaagc agtagaactg ggtgctacta
     2161 ttcctgccgc caatggcaac ggaactcatc agtcaattaa ggttcctgga catcgtcaag
     2221 tgactccagg caagactacg attatggcta ttggccgtgc tgtgcccgct aacacgacgt
     2281 tcaatgatgg tttggcagac cattacatcc aggagttcaa tctgcaagat ccagtccttc
     2341 aggctaaact gcgtcgtttg tgcgaaacca caaccgtgaa aacgcgttat ctggtcgtca
     2401 ataaggagat tctggatgag catcctgagt tcctggttga tggagctgca accgtatctc
     2461 agcgtctggc tattacaggc gaagctgtta cccaacttgg acatgaggca gcaacggctg
     2521 ccatcaaaga gtggggccgt ccagcctccg aaattaccca tcttgtttat gtttcctcat
     2581 ccgaaatccg tctgccaggt ggcgacttgt acctggcaca attgcttggc ttgcgttcag
     2641 acgtgaaccg tgtaatgttg tacatgctgg gttgctatgg tggtgcatct ggaatccgtg
     2701 ttgccaaaga cttggctgag aataacccag gctcacgtgt acttctgatc acgtcagaat
     2761 gcactttgat tggatataag tcactttccc ctgaccgtcc ttacgacctg gttggagcag
     2821 ctctgttcgg tgacggtgct gccgctatga ttatgggaaa agatccaatc cctgttttgg
     2881 agcgtgcttt ctttgagctt gactgggccg gtcagtcttt tattcctgga actaacaaaa
     2941 ccatcgatgg tcgtttgtct gaagaaggca tttccttcaa gctgggtcgt gaactgccta
     3001 agttgattga atccaatatt cagggtttct gcgatcctat cttgaaacgt gctggtggtc
     3061 tgaagtataa cgatattttc tgggcagttc acccaggcgg accagctatc ttgaatgctg
     3121 tccagaaaca actggatttg gcacccgaga aacttcagac cgcacgtcag gttctgcgtg
     3181 actacggtaa catttctagt tccacctgca tttacgtact ggactatatg cgtcatcaat
     3241 ccttgaaact gaaggaagca aatgacaacg taaatacgga accagagtgg ggtctgctgc
     3301 ttgcatttgg ccccggcgtc actatcgagg gtgctttgct gcgtaacctg tgctcaggta
     3361 gttaactagc ttaataggtc ctgctttaat gagatatgcg agaagcctat gatcgcatga
     3421 tatttgcttt caattctgtt gtgcacgttg taaaaaacct gagcatgtgt agctcagatc
     3481 cttaccgccg gtttcggttc attctaatga atatatcacc cgttactatc gtatttttat
     3541 gaataatatt ctccgttcaa tttactgatt gtaccctact acttatatgt acaatattaa
     3601 aatgaaaaca atatattgtg ctgaataggt ttatagcgac atctatgata gagcgccaca
     3661 ataacaaaca attgcgtttt attattacaa atccaatttt aaaaaaagcg gcagaaccgg
     3721 tcaaacctaa aagactgatt acataaatct tattcaaatt tcaaaagtgc cccaggggct
     3781 agtatctacg acacaccgag cggcgaacta ataacgctca ctgaagggaa ctccggttcc
     3841 ccgccggcgc gcatgggtga gattccttga agttgagtat tggccgtccg ctctaccgaa
     3901 agttacgggc accattcaac ccggtccagc acggcggccg ggtaaccgac ttgctgcccc
     3961 gagaattatg cagcattttt ttggtgtatg tgggccccaa atgaagtgca ggtcaaacct
     4021 tgacagtgac gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa catgtcgagg
     4081 ctcagcagga ccgcttgcct tgtcttctgc acgaagtggt ttaaactatc agtgtttgac
     4141 aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt
     4201 aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt
     4261 tccccagatc aggcgctggc tgctgaaccc ccagccggaa ctgaccccac aaggccctag
     4321 cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc
     4381 aactcttcgc aggcttcgcc gacctgctcg cgccacttct tcacgcgggt ggaatccgat
     4441 ccgcacatga ggcggaaggt ttccagcttg agcgggtacg gctcccggtg cgagctgaaa
     4501 tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt acttctccca tatgaatttc
     4561 gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt cgatcaggac ctggcaacgg
     4621 gacgttttct tgccacggtc caggacgcgg aagcggtgca gcagcgacac cgattccagg
     4681 tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga caggcattcc
     4741 tcggccttcg tgtaataccg gccattgatc gaccagccca ggtcctggca aagctcgtag
     4801 aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg cgtactccaa cacctgctgc
     4861 cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc
     4921 ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc
     4981 gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac
     5041 ggcgcaatat cgaacaagga aagctgcatt tccttgatct gctgcttcgt gtgtttcagc
     5101 aacgcggcct gcttggcctc gctgacctgt tttgccaggt cctcgccggc ggtttttcgc
     5161 ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc
     5221 tcctgttcaa gacgacgcga acgctccacg gcggccgatg gcgcgggcag ggcaggggga
     5281 gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt gctggaccat cgagccgacg
     5341 gactggaagg tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc
     5401 tcggcggaaa accccgcgtc gatcagttct tgcctgtatg ccttccggtc aaacgtccga
     5461 ttcattcacc ctccttgcgg gattgccccg actcacgccg gggcaatgtg cccttattcc
     5521 tgatttgacc cgcctggtgc cttggtgtcc agataatcca ccttatcggc aatgaagtcg
     5581 gtcccgtaga ccgtctggcc gtccttctcg tacttggtat tccgaatctt gccctgcacg
     5641 aataccagcg accccttgcc caaatacttg ccgtgggcct cggcctgaga gccaaaacac
     5701 ttgatgcgga agaagtcggt gcgctcctgc ttgtcgccgg catcgttgcg ccacatctag
     5761 gatctgccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc tcaccggctc
     6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg c
//
