LOCUS       sequence-309733-        5379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for Golden Gate assembly. Golden gate compatible N. 
            benthamiana ZAR1 D481V/L17E without STOP codon .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5379)
  AUTHORS   Harant A, Pai H, Sakai T, Kamoun S, Adachi H
  TITLE     A vector system for fast-forward studies of the HOPZ-ACTIVATED 
            RESISTANCE1 (ZAR1) resistosome in the model plant Nicotiana 
            benthamiana.
  JOURNAL   Plant Physiol. 2022 Jan 20;188(1):70-80. doi: 
            10.1093/plphys/kiab471.
  PUBMED    34633454
REFERENCE   2  (bases 1 to 5379)
  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"; journalName: 
            "Plant Physiol"; date: "2022-01-20- 20"; volume: "188"; issue: "1"; 
            pages: "70-80"
COMMENT     Sequence Label: pZA16
FEATURES             Location/Qualifiers
     source          1..5379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2324..2335)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3258..3357)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3374..3393)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3375..3393)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3398..3414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3398..3414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3845..4636
                     /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"
     rep_origin      4729..5317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5218..5237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcgccc ttaatggtct ctcgaaccgt tcctatgttc ttccttcttc
      721 catagacccc ctctaaaccc tgcatcttca attggaaatg actccaactc agggcaccag
      781 ctaaccttga gtattcttaa cgaaggcatg acttgatgca ttgctgacca ttccaatctc
      841 aaatccgata aagattccaa cattaagccc tcaattttcc aagcagtact accatcaccc
      901 cagaagttct cgtgcatttg tgtaatatca ccagagatga ttgataaata tcgcaacatg
      961 ggggagagat tggggttaag ccattcagga cttgtcttac ttggataaaa tttgagaatc
     1021 agctcgtgga gttgtcgtgg aggataaagt ctaccaagtt ttgtcgcaag tccattatct
     1081 tgactgccaa agcaacttat tgttaaaaat tgaagttctt ggagatctac cagtgcgttc
     1141 ccctcatcgt ctccaatctc ctcattttcg gttagtctta aactgagagt tctgagtcga
     1201 gtcaggcttc tgagctcagc aatacgacaa cctcctggtt ggcttaattt tgcaggctta
     1261 aacccaagca atacttgaag attggaaagc ctacttaatc cttttggaag gtactcaagg
     1321 gaaccacagt tgttcacatc caaaacagca agttcctcaa atgtcagaac gcaagagggg
     1381 agcattttca tattttggca atagctgaaa tccaaaatct gtaatttttc aagcttggat
     1441 atggaagctg gaactccaat cattggatgt gtattgctta aactgagata agtgagctgc
     1501 ttagcagatc caattccttc caacaaactt gaaagaggca catcgaaaat tgattttgac
     1561 agatccaata cttggaggtg tcgactcttg cacagcttct tggcaatgtc ggaatggatt
     1621 ttgttaactt caccactctt ggtggttgtt agcagtgctc gtaatttttg attgctcaat
     1681 agttgcttcc aattcatctc actcttaata cccaaatgcc gacaagttgc atcagctggg
     1741 gtgaaaaatg catcatcctc ggctaatttg atcaccaagt cacgaaccat aacatgcatt
     1801 ttgcatgtgt ggatcacacc attgtaagcc ttatcaacaa cttctatcaa acaccgattg
     1861 gatagttgtg agaaacaatc ttctcccact tcagtcgata acctaccact tctcagaggt
     1921 atgaaacctt ctccaatcca ccaacggatc agttggtctt ttggtatgac acaatcctca
     1981 ggaaagagtg aaaaacagag gaaacaggat ttcaagtaag gagggagttc atcatagctc
     2041 aactgtagtg aggccatcac tgagtcatca ttttctttca attcatcacg gaaatgattt
     2101 gaaatacgcc tccattcgtg atagtaagat ggcttgcaaa gcatcactcc tcctactgct
     2161 ttgattgcta aaggaagtcc cttgcatttt tccacaatct cctttcccac gtcctccaac
     2221 tcggggaaat ggcattcacc tgcacttcct gcaaatgcaa tcttccggaa tagtaaccaa
     2281 ctgtagtgct cattgaggaa ttttggccaa tgtattcttg cttctgtaac tcccatcttg
     2341 cgagcgacta gctcatttct cgttgttaca ataacagtgc tcccatttcc tttgggcagt
     2401 ccagtataga ttttttgcca ccaggcattg tcccagctcc aaacatcatc cataacaatc
     2461 aaaaacctct ttcccaaaag gtactggttt atttttctta acagttcaca ctggtcatca
     2521 ccaacacatg catctcctaa actcctcaat atgcttctca tgacttgttc ctcagtaaat
     2581 gtttgagaaa cagacaccca aatcctcctc tccaagtgat cctccaccct tttgtcattg
     2641 aaaactttct gagcaagagt ggttttacca agtcctccca tacccacaaa tgcaatggta
     2701 agtaaaccat ctcttgcttc aaatagccaa tcctttatct tctcagtgtc accttccaaa
     2761 ccaactacct gagtgtggtc ataaagggaa gaagtccatc ttgacattag attattgtgt
     2821 gcctccatac ttccttcttc gagaagtggt actccaagat atgttgaaat gttttgcttt
     2881 atctccgaga ttctgtcatt gattttagca aggcacttcc cagtttgatg gcgaagagag
     2941 agccttttag gatggaagca tgtagtagcc cgatcgctat ctgcagattg attctggcag
     3001 tcctcaagta tctcttcagc ttcaaagatt aagtctcgca aacaggacat gactccttta
     3061 agggtgttgt ttttcctctt gagcctctct gcatctttga gaaagctttg catgaatagc
     3121 agctcgttct tcagtttttc aaactgctgt ctgtatttag taagaaacct actctcctct
     3181 gtttcaacat gcagaagttt ctccaagaat acagtgacca ccgcatccac cattagagac
     3241 cattaagggc gaattcgacc cagctttctt gtacaaagtt ggcattataa aaaataattg
     3301 ctcatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3361 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3421 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     3481 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     3541 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     3601 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     3661 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     3721 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     3781 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     3841 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     3901 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     3961 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     4021 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     4081 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     4141 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     4201 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     4261 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     4321 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     4381 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     4441 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     4501 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     4561 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     4621 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     4681 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4741 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4801 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4861 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5341 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
