LOCUS       sequence-438202-        8858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo-compatible Level M vector encoding hygromycin resistance 
            cassette and a luciferase gene under control of p35S promoter and 
            act2 terminator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8858)
  AUTHORS   Balakireva AV, Karataeva TA, Karampelias M, Mitiouchkina TY, 
            Machacek J, Shakhova ES, Perfilov MM, Belozerova OA, Palkina KA, 
            Drazna N, Vondrakova Z, Fleiss A, Fakhranurova LI, Markina NM, 
            Morozov VV, Bugaeva EN, Delnova GM, Choob VV, Yampolsky IV, Petrasek
            J, Mishin AS, Sarkisyan KS
  TITLE     Non-invasive imaging of salicylic and jasmonic acid activities in 
            planta
  JOURNAL   bioRxiv 2025.02.17.636591; doi: 
            https://doi.org/10.1101/2025.02.17.636591
REFERENCE   2  (bases 1 to 8858)
  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: "bioRxiv 
            2025.02.17.636591; doi: https://doi.org/10.1101/2025.02.17.636591"
COMMENT     Sequence Label: pNK3558
FEATURES             Location/Qualifiers
     source          1..8858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          259..448
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     misc_feature    789..844
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(880..1059)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    1233..1257
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(1403..2551)
                     /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(2887..3475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2967..2986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3816..4607)
                     /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      4902..5612
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5700..5724
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        5872..6217
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     6160..6180
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(6778..6789)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(7715..8422)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
ORIGIN
        1 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
       61 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
      121 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
      181 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
      241 catggcctcc gcgaccggct gcagttatca tcatcatcat agacacacga aataaagtaa
      301 tcagattatc agttaaagct atgtaatatt tacaccataa ccaatcaatt aaaaaataga
      361 tcagtttaaa gaaagatcaa agctcaaaaa aataaaaaga gaaaagggtc ctaaccaaga
      421 aaatgaagga gaaaaactag aaatttacct gcagaacagc gggcagttcg gtttcaggca
      481 ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga
      541 attccccaat gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa
      601 cataacgatc tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc
      661 ctcctacatc gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg
      721 agacactgtc gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct
      781 ttttcattgt aattgtaaat agtaattgta atgttgtttg ttgtttgttg ttgttggtaa
      841 ttgttgtaaa aatacagtat gagagtgaat atgagactct aattggatac cgaggggaat
      901 ttatggaacg tcagtggagc atttttgaca agaaatattt gctagctgat agtgacctta
      961 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     1021 gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     1081 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctctccc
     1141 catgggaatt cgtaatgaga cccacagagt gttcaacccc agtacattaa aaacgtccgc
     1201 aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca
     1261 gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc
     1321 ccatcaacca tgactactgt ccggtccaac agatccgagc aggggctgct gaacccccag
     1381 ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg
     1441 cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc
     1501 acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg
     1561 ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct
     1621 tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt
     1681 cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc
     1741 ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg
     1801 tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc
     1861 agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc
     1921 gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga
     1981 cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg
     2041 cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg
     2101 gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct
     2161 tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg
     2221 ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg
     2281 tcatcgactt cgccaaacct gccgcctcct gttcaagacg acgcgaacgc tccacggcgg
     2341 ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg
     2401 tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg
     2461 tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc
     2521 tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc
     2581 acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat
     2641 aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact
     2701 tggtattccg aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt
     2761 gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt
     2821 cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct
     2881 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2941 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3001 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3061 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3121 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3181 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3241 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3301 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3361 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3421 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagcatga tatatctccc
     3661 aatttgtgta gggcttatta tgcacgctta aaaataataa aagcagactt gacctgatag
     3721 tttggctgtg agcaattatg tgcttagtgc atctaacgct tgagttaagc cgcgccgcga
     3781 agcggcgtcg gcttgaacga atttctagct agacattatt tgccgactac cttggtgatc
     3841 tcgcctttca cgtaatggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     3901 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
     3961 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     4021 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     4081 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     4141 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     4201 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     4261 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     4321 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     4381 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     4441 acggtcaccg taaccagcaa atcaatatcg ctgtgtggct tcaggccgcc atccactgcg
     4501 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     4561 acctctgata gttgagtcga tacttcggcg atcaccgctt cccccatgat gtttaacttt
     4621 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
     4681 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa
     4741 cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag
     4801 gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg aaccgaacag
     4861 gcttatgtcc actgggttcg tgcctcggga attggccagc gctgccattt ttggggtgag
     4921 gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcgggccg
     4981 ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc
     5041 agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt
     5101 agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca
     5161 gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca
     5221 aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac
     5281 ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc
     5341 cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg
     5401 tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag
     5461 tgagggccaa gttttccgcg aggtatccac aacgccggcg gccgcggtgt ctcgcacacg
     5521 gcttcgacgg cgtttctggc gcgtttgcag ggccatagac ggccgccagc ccagcggcga
     5581 gggcaaccag cccggtgagc gtcgcaaagg cggctggcac atacaaatgg acgaacggat
     5641 aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg
     5701 tttacccgcc aatatatcct gtcaaacact gatagtttaa acggatcact tcgtgtccct
     5761 cggtcacatg tgcatcctcg gtctcacaca ggaggtcaac atggtggagc acgacactct
     5821 ggtctactcc aaaaatgtca aagatacagt ctcagaagat caaagggcta ttgagacttt
     5881 tcaacaaagg ataatttcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
     5941 catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg
     6001 aaaggctatc attcaagatc tctctgccga cagtggtccc aaagatggac ccccacccac
     6061 gaggagcatc gtggaaaaag aagaggttcc aaccacgtct acaaagcaag tggattgatg
     6121 tgacatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc
     6181 ctctatataa ggaagttcat ttcatttgga gaggacacgc tcgagtataa gagctcattt
     6241 ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacat ttacaattat
     6301 cgatacaatg aggtctacta tctctttgtc tagtctcctt gagagactga gcaaattgtc
     6361 atcccgatcc atcgcaatta cctgcggcgt agtcttggca agtgcaatag cattcccaat
     6421 tatccgaaga gactatcaaa cgttccttga agtcggtccg agctatgccc cacagaactt
     6481 ccgaggctat atcactgttt gtgtcttgtc actttttagg caagagcaga aagggttggc
     6541 aatctatgac aggcttccag aaaagaggcg ttggcttgcc gacttgccgt ttcgtgaagg
     6601 gccaaggcca tccataacct cccacattat acagcgtcag cgtactcagc tggtagatca
     6661 agagtttgca accagagaac ttatcgataa ggtgatccca cgtgtgcaag caagacacac
     6721 agacaaaact tttctcagta cgtcaaaatt tgagtttcat gcaaaggcca tcttccttct
     6781 cccctctatc cctattaatg atcctcttaa cattccctca cacgatacgg taagaagaac
     6841 caagcgtgag atcgctcaca tgcatgatta ccatgattgc tctctgcact tggctcttgc
     6901 tcctcaggat ggtaaggaag ttttgaagaa ggggtggggc cagcgtcacc cactggccgg
     6961 accaggagtc ccaggccctc ctactgagtg gaccttcctt tacgcaccaa ggaacgaaga
     7021 ggaggccaga gttgtcgaga tgatagtcga agctagtatt ggctacatga ctaacgatcc
     7081 tgctggtaag attgtcgaaa atgccaattc gtccggactc agatctcgag ctcaagcttc
     7141 gaattctgca tgcaagaact ggttctctag cctgtcccac ttcgtcatcc atctgtgagc
     7201 ttgctctcaa gatcaaaggc ttaaaaagct ggggttttat gaatgggatc aaagtttctt
     7261 tttttctttt atatttgctt ctccatttgt ttgtttcatt tccctttttg ttttcgtttc
     7321 tatgatgcac ttgtgtgtga caaactctct gggtttttac ttacgtctgc gtttcaaaaa
     7381 aaaaaaccgc tttcgttttg cgttttagtc ccattgtttt gtagctctga gtgatcgaat
     7441 tgatgcctct ttattccttt tgttccctat aatttctttc aaaactcaga agaaaaacct
     7501 tgaaactctt tgcaatgtta atataagtat tgtataagat ttttattgat ttggttatta
     7561 gtcttacttt tgctacctcc atcttcactt ggaactgata ttctgaatag ttaaagcgtt
     7621 acatgtgttc cattcacaaa tgaacttaaa ctagcacaaa gtcagatatt ttaagatcgc
     7681 accatttcgc tgcatgcgaa ttctctgagc ggtcctgctg agcctcgaca tgttgtcgca
     7741 aaattcgccc tggacccgcc caacgatttg tcgtcactgt caaggtttga cctgcacttc
     7801 atttggggcc cacatacacc aaaaaaatgc tgcataattc tcggggcagc aagtcggtta
     7861 cccggccgcc gtgctggacc gggttgaatg gtgcccgtaa ctttcggtag agcggacggc
     7921 caatactcaa cttcaaggaa tctcacccat gcgcgccggc ggggaaccgg agttcccttc
     7981 agtgagcgtt attagttcgc cgctcggtgt gtcgtagata ctagcccctg gggcactttt
     8041 gaaatttgaa taagatttat gtaatcagtc ttttaggttt gaccggttct gccgcttttt
     8101 ttaaaattgg atttgtaata ataaaacgca attgtttgtt attgtggcgc tctatcatag
     8161 atgtcgctat aaacctattc agcacaatat attgttttca ttttaatatt gtacatataa
     8221 gtagtagggt acaatcagta aattgaacgg agaatattat tcataaaaat acgatagtaa
     8281 cgggtgatat attcattaga atgaaccgaa accggcggta aggatctgag ctacacatgc
     8341 tcaggttttt tacaacgtgc acaacagaat tgaaagcaaa tatcatgcga tcataggctt
     8401 ctcgcatatc tcattaaagc aggacaagcc tattcctttg ccctcggacg agtgctgggg
     8461 cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt
     8521 ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat
     8581 cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg
     8641 tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc
     8701 ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag
     8761 atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct
     8821 gttatgcggc cattgtccgt caggacattg ttggagcc
//
