LOCUS       sequence-472169-        8260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AvrSr62-7 in protoplasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8260)
  AUTHORS   Chen R, Chen J, Outram MA, Powell OR, Arndell T, Gajendiran K, Wang 
            YL, Ayliffe MA, Blundell C, Figueroa M, Sperschneider J, Vanhercke 
            T, Tang D, Xu Y, Zhong G, Gardener C, Yu G, Gourdoupis S, Jaremko Ł,
            Matny O, Steffenson BJ, Boshoff WHP, Meyer WB, Arold ST, Dodds PN, 
            Wulff BBH
  TITLE     A wheat tandem kinase sensor activates an NLR helper to trigger 
            immunity
  JOURNAL   bioRxiv 2024.08.30.610287
REFERENCE   2  (bases 1 to 8260)
  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"; journalName: "bioRxiv 
            2024.08.30.610287"
COMMENT     Sequence Label: pICH47811_AvrSr62-7
FEATURES             Location/Qualifiers
     source          1..8260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    109..133
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      225..856
                     /label=oriV
                     /note="incP origin of replication"
     promoter        943..1047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1048..1116,1117..1908)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1266..1285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2079..2667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2568..2587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3003..4151
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4307..4331
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6400..8052
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(6466..6484)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    join(8087..8260,1)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 cgctaagctt gaattcttgc ttgtcttcac agagtggggc ccactgcatc caccccagta
       61 cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt ttacaccaca
      121 atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca caaaatcacc
      181 actcgataca ggcagcccat cagtcagatc aggatctcct ttgcgacgct caccgggctg
      241 gttgccctcg ccgctgggct ggcggccgtc tatggccctg caaacgcgcc agaaacgccg
      301 tcgaagccgt gtgcgagaca ccgcggccgc cggcgttgtg gatacctcgc ggaaaacttg
      361 gccctcactg acagatgagg ggcggacgtt gacacttgag gggccgactc acccggcgcg
      421 gcgttgacag atgaggggca ggctcgattt cggccggcga cgtggagctg gccagcctcg
      481 caaatcggcg aaaacgcctg attttacgcg agtttcccac agatgatgtg gacaagcctg
      541 gggataagtg ccctgcggta ttgacacttg aggggcgcga ctactgacag atgaggggcg
      601 cgatccttga cacttgaggg gcagagtgct gacagatgag gggcgcacct attgacattt
      661 gaggggctgt ccacaggcag aaaatccagc atttgcaagg gtttccgccc gtttttcggc
      721 caccgctaac ctgtctttta acctgctttt aaaccaatat ttataaacct tgtttttaac
      781 cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc cccttctcga
      841 accctcccgg cccgctaacg cgggcctccc atccccccag gggctgcgcc cctcggccgc
      901 gaacggcctc accccaaaaa tggcagcgct ggccaattcg tgcgcggaac ccctatttgt
      961 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ttgataaatg
     1021 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1081 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1141 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1201 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1261 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1321 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1381 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1441 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1501 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1561 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1621 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1681 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1741 aaatctggag ccggtgagcg tggttctcgc ggtatcattg cagcactggg gccagatggt
     1801 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1861 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1921 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2701 gcagatccta gatgtggcgc aacgatgccg gcgacaagca ggagcgcacc gacttcttcc
     2761 gcatcaagtg ttttggctct caggccgagg cccacggcaa gtatttgggc aaggggtcgc
     2821 tggtattcgt gcagggcaag attcggaata ccaagtacga gaaggacggc cagacggtct
     2881 acgggaccga cttcattgcc gataaggtgg attatctgga caccaaggca ccaggcgggt
     2941 caaatcagga ataagggcac attgccccgg cgtgagtcgg ggcaatcccg caaggagggt
     3001 gaatgaatcg gacgtttgac cggaaggcat acaggcaaga actgatcgac gcggggtttt
     3061 ccgccgagga tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct
     3121 tccagtccgt cggctcgatg gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc
     3181 aactggctcc ccctgccctg cccgcgccat cggccgccgt ggagcgttcg cgtcgtcttg
     3241 aacaggaggc ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga actatgacga
     3301 ccaagaagcg aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgag gccaagcagg
     3361 ccgcgttgct gaaacacacg aagcagcaga tcaaggaaat gcagctttcc ttgttcgata
     3421 ttgcgccgtg gccggacacg atgcgagcga tgccaaacga cacggcccgc tctgccctgt
     3481 tcaccacgcg caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc attttccacg
     3541 tcaacaagga cgtgaagatc acctacaccg gcgtcgagct gcgggccgac gatgacgaac
     3601 tggtgtggca gcaggtgttg gagtacgcga agcgcacccc tatcggcgag ccgatcacct
     3661 tcacgttcta cgagctttgc caggacctgg gctggtcgat caatggccgg tattacacga
     3721 aggccgagga atgcctgtcg cgcctacagg cgacggcgat gggcttcacg tccgaccgcg
     3781 ttgggcacct ggaatcggtg tcgctgctgc accgcttccg cgtcctggac cgtggcaaga
     3841 aaacgtcccg ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc
     3901 actacacgaa attcatatgg gagaagtacc gcaagctgtc gccgacggcc cgacggatgt
     3961 tcgactattt cagctcgcac cgggagccgt acccgctcaa gctggaaacc ttccgcctca
     4021 tgtgcggatc ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg
     4081 aagagttgcg aggcagcggc ctggtggaac acgcctgggt caatgatgac ctggtgcatt
     4141 gcaaacgcta gggccttgtg gggtcagttc cggctggggg ttcagcagcc agcgcctgat
     4201 ctggggaacc ctgtggttgg catgcacata caaatggacg aacggataaa ccttttcacg
     4261 cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat
     4321 atatcctgtc aaacactgat agtttaaacc acttcgtgca gaagacaata gtggagggtc
     4381 gtgcccctct ctagagataa tgagcattgc atgtctaagt tataaaaaat taccacatat
     4441 tttttttgtc acacttgttt gaagtgcagt ttatctatct ttatacatat atttaaactt
     4501 tactctacga ataatataat ctatagtact acaataatat cagtgtttta gagaatcata
     4561 taaatgaaca gttagacatg gtctaaagga caattgagta ttttgacaac aggactctac
     4621 agttttatct ttttagtgtg catgtgttct cctttttttt tgcaaatagc ttcacctata
     4681 taatacttca tccattttat tagtacatcc atttagggtt tagggttaat ggtttttata
     4741 gactaatttt tttagtacat ctattttatt ctattttagc ctctaaatta agaaaactaa
     4801 aactctattt tagttttttt atttaataat ttagatataa aatagaataa aataaagtga
     4861 ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca tttttcttgt
     4921 ttcgagtaga taatgccagc ctgttaaacg ccgtcgatcg acgagtctaa cggacaccaa
     4981 ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtcg
     5041 ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca
     5101 tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc
     5161 ctctcacggc accggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc
     5221 ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt
     5281 tcggagcgca cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt
     5341 caaggtacgc cgctcgtcct cccccccccc ccctctctac cttctctaga tcggcgttcc
     5401 ggtccatggt tagggcccgg tagttctact tctgttcatg tttgtgttag atccgtgttt
     5461 gtgttagatc cgtgctgcta gcgttcgtac acggatgcga cctgtacgtc agacacgttc
     5521 tgattgctaa cttgccagtg tttctctttg gggaatcctg ggatggctct agccgttccg
     5581 cagacgggat cgatctagga taggtataca tgttgatgtg ggttttactg atgcatatac
     5641 atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat ctattataat
     5701 aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg catatgcagc
     5761 agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct tggtactgtt
     5821 tcttttgtcg atgctcaccc tgttgtttgg tgttacttct gcagaatgta cgcgtcccgg
     5881 ggcggtacca tgaagccaat catgaagtcc tccatcttca agggcggcga gactagctct
     5941 ctcggcgagt ctccattcga gggcccactc tacaagtacg aggaaatcgt gcacgcccaa
     6001 ggcaagttcc tcgtgaccac ctactacagg accccaatca agctcgagtt cccattcaac
     6061 ggcaaggaca tctccatcag cctcaacgag ttctccaaga actccgacag cctcggcatc
     6121 tccaaggaag agaaggaaaa ggtcaccaag accatcaact ccgaaggcaa cggcgcccaa
     6181 tcctccacca tgggccaaaa caagggcaag gaactcagca tcgaggaaaa ggaaaagatc
     6241 tggccggacg gcccaccaag gttcgacgag aagaagtacg gcatcaagct cagggacgcc
     6301 aacggcgaca ggaccatctt ttacaagtac tctcacaccc catacgtggt gaccgagctg
     6361 aacttgaacc cagtcgacct gcagagcttt cgtggttcga tggaggacgc caaaaacata
     6421 aagaaaggcc cggcgccatt ctatcctcta gaggatggaa ccgctggaga gcaactgcat
     6481 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     6541 gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc agaagctatg
     6601 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     6661 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     6721 atttataatg aacgtgaatt gctcaacagt atgaacattt cgcagcctac cgtagtgttt
     6781 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaaattacc aataatccag
     6841 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     6901 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtaccaga gtcctttgat
     6961 cgtgacaaaa caattgcact gataatgaat tcctctggat ctactgggtt acctaagggt
     7021 gtggcccttc cgcatagaac tgcctgcgtc agattctcgc atgccagaga tcctattttt
     7081 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     7141 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     7201 tttgaagaag agctgttttt acgatccctt caggattaca aaattcaaag tgcgttgcta
     7261 gtaccaaccc tattttcatt cttcgccaaa agcactctga ttgacaaata cgatttatct
     7321 aatttacacg aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt
     7381 gcaaaacgct tccatcttcc agggatacga caaggatatg ggctcactga gactacatca
     7441 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     7501 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcagaga
     7561 ggcgaattat gtgtcagagg acctatgatt atgtccggtt atgtaaacaa tccggaagcg
     7621 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     7681 gaggacgaac acttcttcat agttgaccgc ttgaagtctt taattaaata caaaggatat
     7741 caggtggccc ccgctgaatt ggaatcgata ttgttacaac accccaacat cttcgacgcg
     7801 ggcgtggcag gtctgcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     7861 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     7921 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     7981 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     8041 tccaaattgt aagcttctct agctagagtc gatcgacaag ctcgagtttc tccataataa
     8101 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
     8161 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
     8221 tctaattcct aaaaccaaaa tccagtacta aaatccagat
//
