LOCUS       sequence-310749-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for Golden Gate assembly. Golden gate compatible N. 
            benthamiana ZAR1 L17E/D481V with STOP codon .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  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 5380)
  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: pZA8
FEATURES             Location/Qualifiers
     source          1..5380
                     /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             1592..1603
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3259..3358)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3375..3394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3376..3394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3399..3415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3399..3415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3846..4637
                     /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      4730..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5219..5238
                     /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 ctaatggtgg atgcggtggt cactgtattc
      721 ttggagaaac ttctgcatgt tgaaacagag gagagtaggt ttcttactaa atacagacag
      781 cagtttgaaa aactgaagaa cgagctgcta ttcatgcaaa gctttctcaa agatgcagag
      841 aggctcaaga ggaaaaacaa cacccttaaa ggagtcatgt cctgtttgcg agacttaatc
      901 tttgaagctg aagagatact tgaggactgc cagaatcaat ctgcagatag cgatcgggct
      961 actacatgct tccatcctaa aaggctctct cttcgccatc aaactgggaa gtgccttgct
     1021 aaaatcaatg acagaatctc ggagataaag caaaacattt caacatatct tggagtacca
     1081 cttctcgaag aaggaagtat ggaggcacac aataatctaa tgtcaagatg gacttcttcc
     1141 ctttatgacc acactcaggt agttggtttg gaaggtgaca ctgagaagat aaaggattgg
     1201 ctatttgaag caagagatgg tttacttacc attgcatttg tgggtatggg aggacttggt
     1261 aaaaccactc ttgctcagaa agttttcaat gacaaaaggg tggaggatca cttggagagg
     1321 aggatttggg tgtctgtttc tcaaacattt actgaggaac aagtcatgag aagcatattg
     1381 aggagtttag gagatgcatg tgttggtgat gaccagtgtg aactgttaag aaaaataaac
     1441 cagtaccttt tgggaaagag gtttttgatt gttatggatg atgtttggag ctgggacaat
     1501 gcctggtggc aaaaaatcta tactggactg cccaaaggaa atgggagcac tgttattgta
     1561 acaacgagaa atgagctagt cgctcgcaag atgggagtta cagaagcaag aatacattgg
     1621 ccaaaattcc tcaatgagca ctacagttgg ttactattcc ggaagattgc atttgcagga
     1681 agtgcaggtg aatgccattt ccccgagttg gaggacgtgg gaaaggagat tgtggaaaaa
     1741 tgcaagggac ttcctttagc aatcaaagca gtaggaggag tgatgctttg caagccatct
     1801 tactatcacg aatggaggcg tatttcaaat catttccgtg atgaattgaa agaaaatgat
     1861 gactcagtga tggcctcact acagttgagc tatgatgaac tccctcctta cttgaaatcc
     1921 tgtttcctct gtttttcact ctttcctgag gattgtgtca taccaaaaga ccaactgatc
     1981 cgttggtgga ttggagaagg tttcatacct ctgagaagtg gtaggttatc gactgaagtg
     2041 ggagaagatt gtttctcaca actatccaat cggtgtttga tagaagttgt tgataaggct
     2101 tacaatggtg tgatccacac atgcaaaatg catgttatgg ttcgtgactt ggtgatcaaa
     2161 ttagccgagg atgatgcatt tttcacccca gctgatgcaa cttgtcggca tttgggtatt
     2221 aagagtgaga tgaattggaa gcaactattg agcaatcaaa aattacgagc actgctaaca
     2281 accaccaaga gtggtgaagt taacaaaatc cattccgaca ttgccaagaa gctgtgcaag
     2341 agtcgacacc tccaagtatt ggatctgtca aaatcaattt tcgatgtgcc tctttcaagt
     2401 ttgttggaag gaattggatc tgctaagcag ctcacttatc tcagtttaag caatacacat
     2461 ccaatgattg gagttccagc ttccatatcc aagcttgaaa aattacagat tttggatttc
     2521 agctattgcc aaaatatgaa aatgctcccc tcttgcgttc tgacatttga ggaacttgct
     2581 gttttggatg tgaacaactg tggttccctt gagtaccttc caaaaggatt aagtaggctt
     2641 tccaatcttc aagtattgct tgggtttaag cctgcaaaat taagccaacc aggaggttgt
     2701 cgtattgctg agctcagaag cctgactcga ctcagaactc tcagtttaag actaaccgaa
     2761 aatgaggaga ttggagacga tgaggggaac gcactggtag atctccaaga acttcaattt
     2821 ttaacaataa gttgctttgg cagtcaagat aatggacttg cgacaaaact tggtagactt
     2881 tatcctccac gacaactcca cgagctgatt ctcaaatttt atccaagtaa gacaagtcct
     2941 gaatggctta accccaatct ctcccccatg ttgcgatatt tatcaatcat ctctggtgat
     3001 attacacaaa tgcacgagaa cttctggggt gatggtagta ctgcttggaa aattgagggc
     3061 ttaatgttgg aatctttatc ggatttgaga ttggaatggt cagcaatgca tcaagtcatg
     3121 ccttcgttaa gaatactcaa ggttagctgg tgccctgagt tggagtcatt tccaattgaa
     3181 gatgcagggt ttagaggggg tctatggaag aaggaagaac ataggaacta agcttagaga
     3241 ccattaaggg cgaattcgac ccagctttct tgtacaaagt tggcattata aaaaataatt
     3301 gctcatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     3361 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     3421 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     3481 gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
     3541 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     3601 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     3661 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     3721 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     3781 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     3841 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     3901 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     3961 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     4021 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     4081 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     4141 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     4201 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     4261 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     4321 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     4381 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     4441 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     4501 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     4561 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     4621 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     4681 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5341 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
