LOCUS       sequence-472166-        8532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-Sr62TK in planta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8532)
  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 8532)
  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_HA-Sr62TK
FEATURES             Location/Qualifiers
     source          1..8532
                     /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"
     promoter        5366..5711
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5654..5674
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5806..5895
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGLYPYDVPDYA"
     CDS             5908..5997
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGLYPYDVPDYA"
     terminator      8278..8530
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 cgctgagctc gaattctagt 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 gaagacaagt aaggaggaat
     4381 tccaatccca caaaaatctg agcttaacag cacagttgct cctctcagag cagaatcggg
     4441 tattcaacac cctcatatca actactacgt tgtgtataac ggtccacatg ccggtatata
     4501 cgatgactgg ggttgtacaa aggcggcaac aaacggcgtt cccggagttg cacacaagaa
     4561 atttgccact attacagagg caagagcagc agctgacgcg tacacaacaa gtcagcaaac
     4621 agacaggttg aacttcatcc ccaaaggaga agctcaactc aagcccaaga gctttgctaa
     4681 ggccctaaca agcccaccaa agcaaaaagc ccactggctc acgctaggaa ccaaaaggcc
     4741 cagcagtgat ccagccccaa aagagatctc ctttgccccg gagattacaa tggacgattt
     4801 cctctatctt tacgatctag gaaggaagtt cgaaggtgaa ggtgacgaca ctatgttcac
     4861 cactgataat gagaaggtta gcctcttcaa tttcagaaag aatgctgacc cacagatggt
     4921 tagagaggcc tacgcagcaa gtctcatcaa gacgatctac ccgagtaaca atctccagga
     4981 gatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaattgcat
     5041 caagaacaca gagaaagaca tatttctcaa gatcagaagt actattccag tatggacgat
     5101 tcaaggcttg cttcataaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt
     5161 tcctactgaa tctaaggcca tgcatggagt ctaagattca aatcgaggat ctaacagaac
     5221 tcgccgtcaa gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga
     5281 aaatcttcgt caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata
     5341 cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc
     5401 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag
     5461 gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatctctctg
     5521 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagagg
     5581 ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac gtaagggatg
     5641 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     5701 tggagaggac acgctcgagt ataagagctc atttttacaa caattaccaa caacaacaaa
     5761 caacaaacaa cattacaatt acatttacaa ttatcgatac ccatgtaccc atacgatgtt
     5821 cctgactatg cgggctatcc ctatgacgtc ccggactatg caggattgta tccatatgac
     5881 gttccagatt acgccactag agctgcttac ccatacgatg ttcctgacta tgcgggctat
     5941 ccctatgacg tcccggacta tgcaggattg tatccatatg acgttccaga ttacgcaatg
     6001 tacgcgtccc ggggcggtac catggacgga tacgagttcg agagggcgga gatagatgca
     6061 ctggagcgcg tcgtacggga tccaagagaa aagccaataa gcctgtcgtt ttggcttctc
     6121 aggcgcataa cgaatgattt ctctaacgca tctgaaattg gtcgaggtgg attttcgaac
     6181 gtttacctgg gggtgcttcc aagtgggttg cgtattgctg tcaaaaagct ttaccaactt
     6241 actgcattgg ctaatgaatt tgaagatgaa gttcttacca caatgagggt tgctcacaaa
     6301 aacatagtgc gactcatagg ctactgtcgt cacacgcaat ctgaagtctt cgaattaaaa
     6361 ggagaatcta ttttcgcaca ggtcagagaa aggttgatct gtacagagta tgtgcctaat
     6421 ggaactcttg atggacatat cactgataag tttcatggag gacttgattg ggaccagcgt
     6481 tatcaagttc tcaaaggaat ttgtcacggt ttgcattatc tgcacgatga aatacgcgtt
     6541 atccacagag atatcaaaac agaaaatata ttgctagatg ataacggcgt gccaaaagtt
     6601 tttgactttg gtttttcgaa gtgccttcct gatggagaat ctttaattaa agatgaagca
     6661 acccagggaa ccctagggta tttggcgccg gagtcagttg ataagcaaga atattccttt
     6721 aagtccgaca tatacagctt tggtgttgtg atcatacaat tattgactgg atcgtggatc
     6781 agtaagaacc gaaatgttga gctagaactt aaggagttga gaaaaaggtt ggtaagagaa
     6841 ggagcatttc catcatggga aaacaaatac caccaagtta gaacatgtct ggagattgtg
     6901 tataactgca tggacatcga cccagataaa aggccgactg cattggaaat tatccaacgg
     6961 cttgatgaaa ccgaaagcgc gaactattct gttagcagtg gccatgcaac accactttgg
     7021 cagtcaggag atgaagaatc cgactcagcg gaagtggatg cttttgagcc agagagaaca
     7081 ccatcaaaag acattctttc agatgatgag gaacccgcat cagcagacgc aaatggagaa
     7141 acgagcacac aaaaacataa tatgcctgat cttgtaagta aacagtcagt gtcggcggaa
     7201 ctgtcctcgc tagatttcct tgagaaaatc acagatggtt tttcatacga gcaaatactc
     7261 gggaaggaaa gtctggacgc agatgttcat aaagcatttc tttatgaggg cagcatttca
     7321 ggtccagcaa aggtagttgt gaagaggtta attggagtag caattccagt tgcgaagttt
     7381 aagagggatg cagaacagct gatgagcctg gatcacaaga atatagtaaa gctccttggc
     7441 tactgccacg acgaatctag agggcataag ctggtacagt ttaaagcaaa accaccgcaa
     7501 gattttaaag gagccgagca acttctctgc tatgaatata tgcacaatgg aagcctccgt
     7561 gagtatctta ctggacaagg atctcgtgaa gttgattggc acatgtgcta caaattgatc
     7621 aaagggacct gcgaaggttt acgctacctc catgagggtt gtggagatcg tccaattctt
     7681 catttgaatc taaacccgtc aaatatactg ctggacgaaa acaacatacc acgcatcacg
     7741 ggttttgatt tttcgaagct cattggtgaa aagaacacaa aatccgtggt tcttaagaag
     7801 aatggaccac taggttacct gccaccggac ttcctgtact caaagggtac tgacctcaag
     7861 tatctaccta cggtggacat atacagcttg ggtcttataa ttttagagat tgcaacacgt
     7921 caagagatca agggagacca tggaataatt attaggactg tcgagacaaa ctggaggcaa
     7981 gatacacaaa tagcaacact gtatggttca ctagaagata aattacggga gcaagtaaaa
     8041 atgtgcattg acattggcct agactgtgtc aagtcaaacc ctaaaaagag acctacagct
     8101 ggggccatca tgctctggct tgacaaaggg agcaaaccgg gagttgccag acctccactc
     8161 cgcgataatg tcacccatgc agatgatcac atccaagaaa aggagaagca gacgccattg
     8221 aggagatttt tcagaagaaa gtaagtcgac ctgcagagct ttcgtgcttg tcaagcagat
     8281 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     8341 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     8401 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     8461 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     8521 ttactagatc ga
//
