LOCUS       sequence-309699-        5400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for Golden Gate assembly. Golden gate compatible A. 
            thaliana ZAR1 D489V/L17E without STOP codon .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5400)
  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 5400)
  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: pZA12
FEATURES             Location/Qualifiers
     source          1..5400
                     /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(3279..3378)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3395..3414)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3396..3414)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3419..3435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3419..3435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3866..4657
                     /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      4750..5338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5239..5258
                     /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 ctcgaaccgg ttctgtgcaa tggtgttttc
      721 atccacactc cgcctctaaa gccaacatct tcaatcgcga aagactctag ctctggacac
      781 cagtttgcgg tcacggtcct caggtacggc attgactgtt gcagcacttc ccagtccatg
      841 tctaggtccg acaatgaact taacattaaa ccttcaattc tccagtgagt attctcgttt
      901 ccccagaatg gttcctgcat tttcaccaaa ttacctgaac atattgacat gtacctcaac
      961 atcggaagct tgtgaggact caaccacgat ggactcgact ttccaggata gaattgtaga
     1021 gaaagctcat ggagctggtg tggaggagta agggcgtcta ttttggttat gagatcatcg
     1081 ccgtagctat catagcagtt gatcgaaatg gacatcagct tgcttaggtt tatcaaagaa
     1141 tcaagctcct cctcttctat ctgatcgcca cgagtcagag aaagaccaag tttccgaagg
     1201 tttgtgagat ttttcacttc actgagcttg cacccattgt tggacctggc tggcttaaat
     1261 cccaacagaa cttcgagctt cacgagactt ccgataccct tggggaagca ttcaagggat
     1321 ccacagttcg tcatgtcaag tacaaggagt ttcttgaaca agacgatgca aggttgaagt
     1381 tgtttcagat tctgacagta gcttgcgtct agaatctgaa ggttgtggag atcttccatg
     1441 gaacgaggaa actggatcaa cggatgtgtg ttgctcaaac taagacatgc taagtgttga
     1501 aggcttgcaa tctcatccaa aatctcagac agaggagcat caaatatgga tttcgagata
     1561 tcgagaaccc gaagatattt gcagtctgtg aatttcttag ctaagtctga gttgagcttg
     1621 ttcacttcac ctgtcttagt tgttgacact acgcctctca gcttatggtt aactttgatc
     1681 tgcttctcat caaagtttcc actaatccca aggtgacgac agttgaggcc ttccgggtta
     1741 gagaacgaat ccttcttggc tatatcgatc accaaatcac gaaccataac atgaatcttg
     1801 caagtgatga tggttccgct ataggttttg tctacaactt ctattagaca cctgtttgtg
     1861 agcccggaaa agcagtcctc tccagattca gttgcagatc ttccgtttct ccacatgaca
     1921 aaaccttctc cgatccatcc atgtactagt tgttgttttg gtataacaca gtcctctgga
     1981 tagagcgaaa gcgtgagaat gcacgactta aggtgggaag ggagttcatc gtagctcaac
     2041 tgcagagatg acataacatt gtcagtttca gaagtgtttc ctcttagctc atcctgaaaa
     2101 tgctctgcaa ttcgtctcca ttcatgatag acatggtctt tgcacagaag taacccacca
     2161 accgctttga ttgtcaaagg caaacctttg catttcgtta caatctcctt gccaacatcc
     2221 tctagctcgg gacgctcaca agtaccgtca tttgctgcaa atgccacatt acaaaacagc
     2281 agccaactgt tatcaggact aaggagctct ggacggtgag tcttgtcgtc tctcgcttgc
     2341 acccttttcg ctacagattc cgaccttgtg gtcacgataa cgctacctcc ttgtcctcta
     2401 ggcagtcctt ggtagatctt gtcccaccag ctcaaattct tgtcccatac atcgtccata
     2461 actatcaagt accttttccc caaaagatac tgctggattt ttctcagcaa tgttccaata
     2521 tcatctccaa cactcgcgtc ccccaagttc ctcaatatgc ttctcatgat ctgttcttca
     2581 gtaaaagtct gggacacaga cacccatatc cttctttcga aacgatgttc gatctctttg
     2641 tcattgaaca cttcttgagc gattgtagtt ttacctagcc ctcccatccc aacaaaggcc
     2701 attatcaaaa gctggctgtc gtttgatcga aagagccact ccttgatttt tcttttatct
     2761 ccttccaaac caaccacctg agtatgatca tacaccggag agctccaccg atcagttcca
     2821 ttatctcttc caacattact cggcgttatg aattcgaaat aaggctctac ttgactcttt
     2881 attttcgtta tcctctcatt gatctcttgc aaacgtttgc ttttcttgta ctgcaacgga
     2941 acccgggcag gatgaagccg cgaaagccac gcatttgatg acctttgctc gttcccatca
     3001 tcaccatccg cgagctgaca atccacaagt atatcctcag cttcgtaaac cagttctcgt
     3061 aaatccgcaa caagagtacg caacgtttca ttagttctct tctgcctctc tgcatccttg
     3121 agaaaagact gcatatattt caattcgctt tgcaaatctt ccaactgttt tcggtagtcc
     3181 gatacggttc ggcctttttc ttcttcgatg ttcaaggttt tctctaaaaa cactgttaca
     3241 acagcgtcca ccattagaga ccattaaggg cgaattcgac ccagctttct tgtacaaagt
     3301 tggcattata aaaaataatt gctcatcaat ttgttgcaac gaacaggtca ctatcagtca
     3361 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     3421 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     3481 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     3541 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     3601 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     3661 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     3721 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     3781 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     3841 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     3901 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     3961 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     4021 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     4081 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     4141 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     4201 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     4261 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     4321 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     4381 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     4441 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     4501 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     4561 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     4621 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     4681 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4741 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4801 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4861 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4921 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4981 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5041 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5101 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5161 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5221 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5281 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5341 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
