LOCUS       sequence-309730-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for Golden Gate assembly. Golden gate compatible N. 
            benthamiana ZAR1 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: pZA5
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             complement(2325..2336)
                     /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 ctaagcttag ttcctatgtt cttccttctt
      721 ccatagaccc cctctaaacc ctgcatcttc aattggaaat gactccaact cagggcacca
      781 gctaaccttg agtattctta acgaaggcat gacttgatgc attgctgacc attccaatct
      841 caaatccgat aaagattcca acattaagcc ctcaattttc caagcagtac taccatcacc
      901 ccagaagttc tcgtgcattt gtgtaatatc accagagatg attgataaat atcgcaacat
      961 gggggagaga ttggggttaa gccattcagg acttgtctta cttggataaa atttgagaat
     1021 cagctcgtgg agttgtcgtg gaggataaag tctaccaagt tttgtcgcaa gtccattatc
     1081 ttgactgcca aagcaactta ttgttaaaaa ttgaagttct tggagatcta ccagtgcgtt
     1141 cccctcatcg tctccaatct cctcattttc ggttagtctt aaactgagag ttctgagtcg
     1201 agtcaggctt ctgagctcag caatacgaca acctcctggt tggcttaatt ttgcaggctt
     1261 aaacccaagc aatacttgaa gattggaaag cctacttaat ccttttggaa ggtactcaag
     1321 ggaaccacag ttgttcacat ccaaaacagc aagttcctca aatgtcagaa cgcaagaggg
     1381 gagcattttc atattttggc aatagctgaa atccaaaatc tgtaattttt caagcttgga
     1441 tatggaagct ggaactccaa tcattggatg tgtattgctt aaactgagat aagtgagctg
     1501 cttagcagat ccaattcctt ccaacaaact tgaaagaggc acatcgaaaa ttgattttga
     1561 cagatccaat acttggaggt gtcgactctt gcacagcttc ttggcaatgt cggaatggat
     1621 tttgttaact tcaccactct tggtggttgt tagcagtgct cgtaattttt gattgctcaa
     1681 tagttgcttc caattcatct cactcttaat acccaaatgc cgacaagttg catcagctgg
     1741 ggtgaaaaat gcatcatcct cggctaattt gatcaccaag tcacgaacca tgtcatgcat
     1801 tttgcatgtg tggatcacac cattgtaagc cttatcaaca acttctatca aacaccgatt
     1861 ggatagttgt gagaaacaat cttctcccac ttcagtcgat aacctaccac ttctcagagg
     1921 tatgaaacct tctccaatcc accaacggat cagttggtct tttggtatga cacaatcctc
     1981 aggaaagagt gaaaaacaga ggaaacagga tttcaagtaa ggagggagtt catcatagct
     2041 caactgtagt gaggccatca ctgagtcatc attttctttc aattcatcac ggaaatgatt
     2101 tgaaatacgc ctccattcgt gatagtaaga tggcttgcaa agcatcactc ctcctactgc
     2161 tttgattgct aaaggaagtc ccttgcattt ttccacaatc tcctttccca cgtcctccaa
     2221 ctcggggaaa tggcattcac ctgcacttcc tgcaaatgca atcttccgga atagtaacca
     2281 actgtagtgc tcattgagga attttggcca atgtattctt gcttctgtaa ctcccatctt
     2341 gcgagcgact agctcatttc tcgttgttac aataacagtg ctcccatttc ctttgggcag
     2401 tccagtatag attttttgcc accaggcatt gtcccagctc caaacatcat ccataacaat
     2461 caaaaacctc tttcccaaaa ggtactggtt tatttttctt aacagttcac actggtcatc
     2521 accaacacat gcatctccta aactcctcaa tatgcttctc atgacttgtt cctcagtaaa
     2581 tgtttgagaa acagacaccc aaatcctcct ctccaagtga tcctccaccc ttttgtcatt
     2641 gaaaactttc tgagcaagag tggttttacc aagtcctccc atacccacaa atgcaatggt
     2701 aagtaaacca tctcttgctt caaatagcca atcctttatc ttctcagtgt caccttccaa
     2761 accaactacc tgagtgtggt cataaaggga agaagtccat cttgacatta gattattgtg
     2821 tgcctccata cttccttctt cgagaagtgg tactccaaga tatgttgaaa tgttttgctt
     2881 tatctccgag attctgtcat tgattttagc aaggcacttc ccagtttgat ggcgaagaga
     2941 gagcctttta ggatggaagc atgtagtagc ccgatcgcta tctgcagatt gattctggca
     3001 gtcctcaagt atctcttcag cttcaaagat taagtctcgc aaacaggaca tgactccttt
     3061 aagggtgttg tttttcctct tgagcctctc tgcatctttg agaaagcttt gcatgaatag
     3121 cagctcgttc ttcagttttt caaactgctg tctgtattta gtaagaaacc tactctcctc
     3181 tgtgagaaca tgcagaagtt tctccaagaa tacagtgacc accgcatcca ccattagaga
     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
//
