LOCUS       sequence-309728-        5401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for Golden Gate assembly. Golden Gate compatible A. 
            thaliana ZAR1 L17E/D489V with STOP codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5401)
  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 5401)
  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: pZA4
FEATURES             Location/Qualifiers
     source          1..5401
                     /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"
     protein_bind    complement(3280..3379)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3396..3415)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3397..3415)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3420..3436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3420..3436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3867..4658
                     /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      4751..5339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5240..5259
                     /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 ctaagcttag gttctgtgca atggtgtttt
      721 catccacact ccgcctctaa agccaacatc ttcaatcgcg aaagactcta gctctggaca
      781 ccagtttgcg gtcacggtcc tcaggtacgg cattgactgt tgcagcactt cccagtccat
      841 gtctaggtcc gacaatgaac ttaacattaa accttcaatt ctccagtgag tattctcgtt
      901 tccccagaat ggttcctgca ttttcaccaa attacctgaa catattgaca tgtacctcaa
      961 catcggaagc ttgtgaggac tcaaccacga tggactcgac tttccaggat agaattgtag
     1021 agaaagctca tggagctggt gtggaggagt aagggcgtct attttggtta tgagatcatc
     1081 gccgtagcta tcatagcagt tgatcgaaat ggacatcagc ttgcttaggt ttatcaaaga
     1141 atcaagctcc tcctcttcta tctgatcgcc acgagtcaga gaaagaccaa gtttccgaag
     1201 gtttgtgaga tttttcactt cactgagctt gcacccattg ttggacctgg ctggcttaaa
     1261 tcccaacaga acttcgagct tcacgagact tccgataccc ttggggaagc attcaaggga
     1321 tccacagttc gtcatgtcaa gtacaaggag tttcttgaac aagacgatgc aaggttgaag
     1381 ttgtttcaga ttctgacagt agcttgcgtc tagaatctga aggttgtgga gatcttccat
     1441 ggaacgagga aactggatca acggatgtgt gttgctcaaa ctaagacatg ctaagtgttg
     1501 aaggcttgca atctcatcca aaatctcaga cagaggagca tcaaatatgg atttcgagat
     1561 atcgagaacc cgaagatatt tgcagtctgt gaatttctta gctaagtctg agttgagctt
     1621 gttcacttca cctgtcttag ttgttgacac tacgcctctc agcttatggt taactttgat
     1681 ctgcttctca tcaaagtttc cactaatccc aaggtgacga cagttgaggc cttccgggtt
     1741 agagaacgaa tccttcttgg ctatatcgat caccaaatca cgaaccataa catgaatctt
     1801 gcaagtgatg atggttccgc tataggtttt gtctacaact tctattagac acctgtttgt
     1861 gagcccggaa aagcagtcct ctccagattc agttgcagat cttccgtttc tccacatgac
     1921 aaaaccttct ccgatccatc catgtactag ttgttgtttt ggtataacac agtcctctgg
     1981 atagagcgaa agcgtgagaa tgcacgactt aaggtgggaa gggagttcat cgtagctcaa
     2041 ctgcagagat gacataacat tgtcagtttc agaagtgttt cctcttagct catcctgaaa
     2101 atgctctgca attcgtctcc attcatgata gacatggtct ttgcacagaa gtaacccacc
     2161 aaccgctttg attgtcaaag gcaaaccttt gcatttcgtt acaatctcct tgccaacatc
     2221 ctctagctcg ggacgctcac aagtaccgtc atttgctgca aatgccacat tacaaaacag
     2281 cagccaactg ttatcaggac taaggagctc tggacggtga gtcttgtcgt ctctcgcttg
     2341 cacccttttc gctacagatt ccgaccttgt ggtcacgata acgctacctc cttgtcctct
     2401 aggcagtcct tggtagatct tgtcccacca gctcaaattc ttgtcccata catcgtccat
     2461 aactatcaag taccttttcc ccaaaagata ctgctggatt tttctcagca atgttccaat
     2521 atcatctcca acactcgcgt cccccaagtt cctcaatatg cttctcatga tctgttcttc
     2581 agtaaaagtc tgggacacag acacccatat ccttctttcg aaacgatgtt cgatctcttt
     2641 gtcattgaac acttcttgag cgattgtagt tttacctagc cctcccatcc caacaaaggc
     2701 cattatcaaa agctggctgt cgtttgatcg aaagagccac tccttgattt ttcttttatc
     2761 tccttccaaa ccaaccacct gagtatgatc atacaccgga gagctccacc gatcagttcc
     2821 attatctctt ccaacattac tcggcgttat gaattcgaaa taaggctcta cttgactctt
     2881 tattttcgtt atcctctcat tgatctcttg caaacgtttg cttttcttgt actgcaacgg
     2941 aacccgggca ggatgaagcc gcgaaagcca cgcatttgat gacctttgct cgttcccatc
     3001 atcaccatcc gcgagctgac aatccacaag tatatcctca gcttcgtaaa ccagttctcg
     3061 taaatccgca acaagagtac gcaacgtttc attagttctc ttctgcctct ctgcatcctt
     3121 gagaaaagac tgcatatatt tcaattcgct ttgcaaatct tccaactgtt ttcggtagtc
     3181 cgatacggtt cggccttttt cttcttcgat gttcaaggtt ttctctaaaa acactgttac
     3241 aacagcgtcc accattagag accattaagg gcgaattcga cccagctttc ttgtacaaag
     3301 ttggcattat aaaaaataat tgctcatcaa tttgttgcaa cgaacaggtc actatcagtc
     3361 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3421 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     3481 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     3541 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3601 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3661 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3721 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3781 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3841 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     3901 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     3961 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     4021 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     4081 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     4141 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     4201 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     4261 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4321 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4381 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4441 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4501 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4561 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4621 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     4681 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5401 t
//
