LOCUS       sequence-472161-        9209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sr62NLRa in planta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9209)
  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 9209)
  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: pICH47822_Sr62NLRa
FEATURES             Location/Qualifiers
     source          1..9209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      57..688
                     /label=oriV
                     /note="incP origin of replication"
     promoter        775..879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(880..948,949..1740)
                     /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(1098..1117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1911..2499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2400..2419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2835..3983
                     /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    4139..4163
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        5198..5543
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5486..5506
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      8787..9039
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    9150..9174
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cacaaaatca ccactcgata caggcagccc atcagtcaga tcaggatctc ctttgcgacg
       61 ctcaccgggc tggttgccct cgccgctggg ctggcggccg tctatggccc tgcaaacgcg
      121 ccagaaacgc cgtcgaagcc gtgtgcgaga caccgcggcc gccggcgttg tggatacctc
      181 gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac
      241 tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc
      301 tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg
      361 tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac
      421 agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac
      481 ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc
      541 ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac
      601 cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc
      661 cccccttctc gaaccctccc ggcccgctaa cgcgggcctc ccatcccccc aggggctgcg
      721 cccctcggcc gcgaacggcc tcaccccaaa aatggcagcg ctggccaatt cgtgcgcgga
      781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      841 ccttgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1081 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1561 tttattgctg ataaatctgg agccggtgag cgtggttctc gcggtatcat tgcagcactg
     1621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2221 tcgggctgaa cgggggggtc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2521 ttacggttcc tggcagatcc tagatgtggc gcaacgatgc cggcgacaag caggagcgca
     2581 ccgacttctt ccgcatcaag tgttttggct ctcaggccga ggcccacggc aagtatttgg
     2641 gcaaggggtc gctggtattc gtgcagggca agattcggaa taccaagtac gagaaggacg
     2701 gccagacggt ctacgggacc gacttcattg ccgataaggt ggattatctg gacaccaagg
     2761 caccaggcgg gtcaaatcag gaataagggc acattgcccc ggcgtgagtc ggggcaatcc
     2821 cgcaaggagg gtgaatgaat cggacgtttg accggaaggc atacaggcaa gaactgatcg
     2881 acgcggggtt ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc atgcgtgcgc
     2941 cccgcgaaac cttccagtcc gtcggctcga tggtccagca agctacggcc aagatcgagc
     3001 gcgacagcgt gcaactggct ccccctgccc tgcccgcgcc atcggccgcc gtggagcgtt
     3061 cgcgtcgtct tgaacaggag gcggcaggtt tggcgaagtc gatgaccatc gacacgcgag
     3121 gaactatgac gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa caggtcagcg
     3181 aggccaagca ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa atgcagcttt
     3241 ccttgttcga tattgcgccg tggccggaca cgatgcgagc gatgccaaac gacacggccc
     3301 gctctgccct gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg caaaacaagg
     3361 tcattttcca cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag ctgcgggccg
     3421 acgatgacga actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc cctatcggcg
     3481 agccgatcac cttcacgttc tacgagcttt gccaggacct gggctggtcg atcaatggcc
     3541 ggtattacac gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg atgggcttca
     3601 cgtccgaccg cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc cgcgtcctgg
     3661 accgtggcaa gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc gtcgtgctgt
     3721 ttgctggcga ccactacacg aaattcatat gggagaagta ccgcaagctg tcgccgacgg
     3781 cccgacggat gttcgactat ttcagctcgc accgggagcc gtacccgctc aagctggaaa
     3841 ccttccgcct catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc gagcaggtcg
     3901 gcgaagcctg cgaagagttg cgaggcagcg gcctggtgga acacgcctgg gtcaatgatg
     3961 acctggtgca ttgcaaacgc tagggccttg tggggtcagt tccggctggg ggttcagcag
     4021 ccagcgcctg atctggggaa ccctgtggtt ggcatgcaca tacaaatgga cgaacggata
     4081 aaccttttca cgccctttta aatatccgat tattctaata aacgctcttt tctcttaggt
     4141 ttacccgcca atatatcctg tcaaacactg atagtttaaa ccacttcgtg cagaagacaa
     4201 gtaaggagga attccaatcc cacaaaaatc tgagcttaac agcacagttg ctcctctcag
     4261 agcagaatcg ggtattcaac accctcatat caactactac gttgtgtata acggtccaca
     4321 tgccggtata tacgatgact ggggttgtac aaaggcggca acaaacggcg ttcccggagt
     4381 tgcacacaag aaatttgcca ctattacaga ggcaagagca gcagctgacg cgtacacaac
     4441 aagtcagcaa acagacaggt tgaacttcat ccccaaagga gaagctcaac tcaagcccaa
     4501 gagctttgct aaggccctaa caagcccacc aaagcaaaaa gcccactggc tcacgctagg
     4561 aaccaaaagg cccagcagtg atccagcccc aaaagagatc tcctttgccc cggagattac
     4621 aatggacgat ttcctctatc tttacgatct aggaaggaag ttcgaaggtg aaggtgacga
     4681 cactatgttc accactgata atgagaaggt tagcctcttc aatttcagaa agaatgctga
     4741 cccacagatg gttagagagg cctacgcagc aagtctcatc aagacgatct acccgagtaa
     4801 caatctccag gagatcaaat accttcccaa gaaggttaaa gatgcagtca aaagattcag
     4861 gactaattgc atcaagaaca cagagaaaga catatttctc aagatcagaa gtactattcc
     4921 agtatggacg attcaaggct tgcttcataa accaaggcaa gtaatagaga ttggagtctc
     4981 taaaaaggta gttcctactg aatctaaggc catgcatgga gtctaagatt caaatcgagg
     5041 atctaacaga actcgccgtc aagactggcg aacagttcat acagagtctt ttacgactca
     5101 atgacaagaa gaaaatcttc gtcaacatgg tggagcacga cactctggtc tactccaaaa
     5161 atgtcaaaga tacagtctca gaagatcaaa gggctattga gacttttcaa caaaggataa
     5221 tttcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag
     5281 tagaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag gctatcattc
     5341 aagatctctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
     5401 aaaaagaaga ggttccaacc acgtctacaa agcaagtgga ttgatgtgac atctccactg
     5461 acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa
     5521 gttcatttca tttggagagg acacgctcga gtataagagc tcatttttac aacaattacc
     5581 aacaacaaca aacaacaaac aacattacaa ttacatttac aattatcgat atggaggaga
     5641 tcgtgcttgg actgtccaag acggtggtgg aggggacgct ggtcaaggtg aaggcggcga
     5701 ttgacgagga gtcgaggctg aagctcgcgg tgcagagcga cctggtgttc atcacggggg
     5761 agttcgagat gatgcagtcg ttcctcaacg tcgccgacgc ggagcgcatc aagaacaacg
     5821 ccgtcaagac gtgggtgagg cagctccgcg acctggccta cgacaccgag gactgcatcg
     5881 agctggtcgt ccacctggat cccaagccca gatggtggcg ccgcctgctg gtcctgcctt
     5941 gcctgccggc ggtgtcgctg ccaatggatg acgcggcggc ggagataaag gagctcaagg
     6001 gcagggtgga gttcgtgagc cagcggaata tgcgctacaa cctcatcacc gacttcggcg
     6061 gcgccaagtc cgccgcgcag cagcagcttg accgggccgc cggagctctc gacatcgccg
     6121 tggaggcggc ggcgaggaaa gggcgtagct tggtggatct gaccgagttg atccccaaaa
     6181 tggaggaccg ccctgagctc ggcgtgatct cggtgtgggg cacaggcggc gaccttgggg
     6241 tggcgtccat cgtcaggaag gcctacgacg accctgaaat acgcaagaac ttccatagtc
     6301 gtggctgggc caagttgacc caccctttcg atccgcggaa gatcctccgg agcttgctca
     6361 ttcagttctg cactaacact gctgctccac agaagcaagg ggagactctg gatgtagatt
     6421 cgttgctgaa gatggaaaag acagtggtgg aagaagggga gctcgtcaag cagtttgtga
     6481 gtcatgtgag cacccacagg ttcctcgtcg tcctcgaagg cgtgtcaacg atggcggaat
     6541 gggatgccct ccggatgtac ctgccagaca tgggtaatgg gagtcagatc atcgtatcca
     6601 cacagcagtt cgacatcgcg agcttgtgca ccgggcaact gcacaaagtt tcagagttga
     6661 gaaaattcac acctgaccac tctgtttgtg tgttcttcaa ggaggtttct ggaggtgatg
     6721 atcctggaaa tgcaatgctg gagggagatg acaaagctga acttattggg cgtgacacag
     6781 aagaggagga gcttatggag cagattgatg aggtttcccg tgtgatctcg gtatggggga
     6841 ttgctggtgt aggaaaatcg gcacttgtca gaagtgccta ctgcaagtat ctccaggacc
     6901 gccccaactt tgctgggtta aatctgacct acatacacgg gcgcgtgacc gtgccccatc
     6961 cctttaatct gagggagttc tgccgcagct tgctcctgaa cttgtcctct gagcctgcta
     7021 aaaccaaaga ggacgccgtc gctgaactgg caaacatgac caatccaact gatgtctgcc
     7081 gtaagcttct tagcgacgat caacaacaga agcagtgcct tattgttatt gatgacctcc
     7141 agtccactga agaatgggac agaataaaac aggccttgtc atttggtaaa gaatgttgta
     7201 tcatagttgt tacgaatgat tcaagtgttg ccaaatattg ttcagggcaa gatcagaaac
     7261 tcatgttcaa cgtcaaaggt cttggtgacg agcatgccct tgacctcttc aacacggtat
     7321 atgacggcat gcctaaccac atcacacggg atgaggacat gatcaatcag tcgaagctaa
     7381 tgttaaacaa atgcggcgga ctacccaagg tgatagttgc aattggtcgt ttcttggcca
     7441 agagtctgaa ttgggaggcc atgaatacta actttattca ccagttggaa aacaacccag
     7501 agctggctag tatacatagc ctatttagtt ggctggactc ctacttccac aactgtcctg
     7561 atgaactcaa gccatgcatc ttctatttgt caatcttccc tcgacgccac ggcattcgtc
     7621 ggaggcgtct ggtgaggcgg tggatcgccg agggctactc aagggacacc gacgggaatc
     7681 tcgcggaagt gaacggggaa aattacttct ccaggcttgt caacctcagc atgctcacag
     7741 atgctgagcg tggtgtcaac gcggctggga cagggaggaa aatggccatg tgcaacatca
     7801 acgatttctt ccgggagtac attgtgtcgc ggcgaatgga ggagaaccat gtgtttgcac
     7861 tggaggggag gtgcagccag accacacggc gcactgggcg gcacctggtc atcaaccaaa
     7921 gctgggacgg agatgagaac gtgttcaaca ggatcgagtt ctcgcgactg cggtcggtga
     7981 cagtgttcgg accctggaag ccattcctgg cctccgacaa gatgagggtg ctccgcgtgc
     8041 ttgatctgga aggcactgaa ggtctgacag acgacgacat caagagcatc gtcaaacggc
     8101 tgcctcgact gaagttcctc tccctgcgaa aatgcaaaaa catctttcgg ctgccgaagt
     8161 ccctgggtcg cctgaggcag ctccagaccc tggatgtgag gcacactgcc atagcccagc
     8221 tgccagcgac cgtcgtgaag ctacagaagt tgcagtatgt tcgtgccggt aacacaccag
     8281 aggaatatgc cggcagcgac agcacggaat ctagcacctc ggtgcccgcc tgtttctcat
     8341 gtggccctgt tggcggtcac attgtcggcg ttgaggtgcc tcgcggggtc gagaatctga
     8401 ccaacctgca cacactgggt gtcatcaaag ccactgtggc gggactcaag gagctcaaaa
     8461 agcttaccca gctgcgcaag ctcggtgtgt ctggcatcaa ccggagaaac cacaaagagc
     8521 tctgtgccgt cgtctccgga catggtcatc tggaatccct gtcgatatgg cttgacaagg
     8581 acaccgaggg gattgctgac ccttgttcgc cgagcaatga tgacttcaag cctccagaga
     8641 agctacggag gctcaaactc cacgggcaca tcggcaattt gccagcatgg gttagcactc
     8701 tcaccatgct ggatttggtg gaagtatcca agtctgctgc tcaagatcca tgagtcgacc
     8761 tgcagagctt tcgtgcttgt caagcagatc gttcaaacat ttggcaataa agtttcttaa
     8821 gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta
     8881 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     8941 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     9001 ataaattatc gcgcgcggtg tcatctatgt tactagatcg acgctgagct cgaattctag
     9061 tttgtcttca cagagtgggg cccactgcat ccaccccagt acattaaaaa cgtccgcaat
     9121 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     9181 agccaacagc tccccgaccg gcagctcgg
//
