LOCUS       sequence-258868-        7621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Blumeria graminis f. sp. hordei effector AVRa1-V1 
            ∆SP from the Zea mays ubiquitin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7621)
  AUTHORS   Saur IML, Bauer S, Lu X, Schulze-Lefert P
  TITLE     A cell death assay in barley and wheat protoplasts for 
            identification and validation of matching pathogen AVR effector and 
            plant NLR immune receptors.
  JOURNAL   Plant Methods. 2019 Oct 24;15:118. doi: 10.1186/s13007-019-0502-0. 
            eCollection 2019.
  PUBMED    31666804
REFERENCE   2  (bases 1 to 7621)
  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: "Plant 
            Methods."; date: "2019-10-24"; volume: "15:118. doi"; pages: " 
            10.1186/s13007-019-0502-0. eCollection 2019"
COMMENT     Sequence Label: pZmUBQ:AVRa1-V1
FEATURES             Location/Qualifiers
     source          1..7621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1950..2129
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2171..2722
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      2742..2994
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3088..3112
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3424..4012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3504..3523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4183..4974,4975..5043))
                     /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     4806..4825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5044..5135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5208..5917
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6029..6053
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(join(6346..7059,7060..7062,7063..7065))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6391..6412)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6740..6759
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6999..7020
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    7073..7097
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7414..7438)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7472..7488)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7473..7489)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 atatcaagat caaaataatt ataaaacata cttgtttatt ataatagata ggtactcaag
       61 gttagagcat atgaatagat gctgcatatg ccatcatgta tatgcatcag taaaacccac
      121 atcaacatgt atacctatcc tagatcgata cttccatcca tcatcttaaa ctcgtaacta
      181 tgaagatgta tggcacacac atacagatcc aaaattaata aatccaccag ttagtttgaa
      241 acagtattct actccgatct agaacgaatg aacgaccgcc caaccacacc acatcatcac
      301 aaccaagcga acaaaaagca tctctgtata tgcatcagta aaacccgcat caacatgtat
      361 acctatccta gatcgatatt tccatccatc atcttcaatt cgtaactatg aatatgtatg
      421 gcacacacat acagatccaa aattaataaa tccaccaggt agtttgaaac agaattctac
      481 tccgatttag aacgaccgcc caaccagacc acatcatcac aaccaagcca aaaaaaaaac
      541 atgaaaagat gacccgacaa acaagtgcac ggcatatatt gaaataaagg aaaagggcaa
      601 accaaaccct atgcaacgaa acaaaaaaaa atcatgaaat cgatcccgtc tgcggaacgg
      661 ctagagccat cccaggattc cccaaagaga aacactggca agttagcaat cagaacgtgt
      721 ctgacgtaca ggtcgcatcc gtgtacgaac gctagcagca cggatctatc acaaacacgg
      781 atctaacaca aacatgaaca gaagtagaac taccgggccc taaccatgga ccggaacgcc
      841 gatctagaga aggtagagag ggggggggga ggacgagcgg cgtaccttga agcggaggtg
      901 ccgacgggtg gatttggggg agatctggtt gtgtgtgtgt gcgctccgaa caacacgagg
      961 ttggggaaag agggtgtgga gggggtgtct atttattacg gcgggcgagg aagggaaagc
     1021 gaaggagcgg tgggaaagga atcccccgta gctgccggtg ccgtgagagg aggaggaggc
     1081 cgcctgccgt gccggctcac gtctgccgct ccgccacgca atttctggat gccgacagcg
     1141 gagcaagtcc aacggtggag cggaactctc gagaggggcc cagaggcagc gacagagatg
     1201 ccgtgccgtc tgcttcgctt ggcccgacgc gacgctgctg gttcgctggt tggtgtccgt
     1261 tagactcgtc gacggcgttt aacaggctgg cattatctac tcgaaacaag aaaaatgttt
     1321 ccttagtttt tttaatttct taaagggtat ttgtttaatt tttagtcact ttattttatt
     1381 ctattttata tctaaattat taaataaaaa aactaaaata gagttttagt tttcttaatt
     1441 tagaggctaa aatagaataa aatagatgta ctaaaaaaat tagtctataa aaaccattaa
     1501 ccctaaaccc taaatggatg tactaataaa atggatgaag tattatatag gtgaagctat
     1561 ttgcaaaaaa aaaggagaac acatgcacac taaagagata aaactgtaga gtcctgttgt
     1621 caaagtactc aattgtcctt tagaccatgt ctaactgttc atttatatga ttctctaaaa
     1681 cactgatatt attgtagtac tatagattat attattcgta gagtgaagtt taaatatatg
     1741 tataaagata gataaactgc acttcaaaca agtgtgacaa aaaaatatgt ggtaattttt
     1801 tataacttag acatgcaatg ctcattaaat cggcgcgccc aaaccgaagg cgggaaacga
     1861 caatctgatc atgagcggag aattaaggga gtcacgttat gacccccgcc gatgacgcgg
     1921 gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc cactgagccg
     1981 cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt gcgcgttcaa
     2041 aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc tccactgacg
     2101 ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa ctcgatcgag
     2161 gggatctacc atgagcccag aacgacgccc ggccgacatc cgccgtgcca ccgaggcgga
     2221 catgccggcg gtctgcacca tcgtcaacca ctacatcgag acaagcacgg tcaacttccg
     2281 taccgagccg caggaaccgc aggagtggac ggacgacctc gtccgtctgc gggagcgcta
     2341 tccctggctc gtcgccgagg tggacggcga ggtcgccggc atcgcctacg cgggtccctg
     2401 gaaggcacgc aacgcctacg actggacggc cgagtcgacc gtgtacgtct ccccccgcca
     2461 ccagcggacg ggactgggct ccacgctcta cacccacctg ctgaagtccc tggaggcaca
     2521 gggcttcaag agcgtggtcg ctgtcatcgg gctgcccaac gacccgagcg tgcgcatgca
     2581 cgaggcgctc ggatatgccc cccgcggcat gctgcgggcg gccggcttca agcacgggaa
     2641 ctggcatgac gtgggtttct ggcagctgga cttcagcctg ccggtgccgc cccgtccggt
     2701 cctgcccgtc accgaaatct gatgacccct agagtcaagc agatcgttca aacatttggc
     2761 aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc
     2821 tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat
     2881 gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat
     2941 agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgaccgg
     3001 catgcaagct gatatctggt agagacgtta gtattcgatc attaaaaacg tccgcaatgt
     3061 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag
     3121 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc
     3181 agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc
     3241 gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg
     3301 agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatcc tctagctaga
     3361 gtccatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5101 atacatattt gaatgtattt agaaaaataa acaaataggg gtagctttcg ggctgatcgc
     5161 ccatcccgtt actgccccga tcccggcaat ggcaaggact gccagcgctg ccatttttgg
     5221 ggtgaggccg ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc
     5281 gggccgggag ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca
     5341 gggcgcagcc ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga
     5401 caggttagcg gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg
     5461 tggacagccc ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa
     5521 gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca
     5581 gggcacttat ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt
     5641 tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct
     5701 catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc
     5761 tgtcagtgag ggccaagttt tccgcgaggt atccacaacg ccggcggccg gtgtctcgca
     5821 cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg
     5881 gcgagggcaa ccagcccggt gagcgtcgga aaggcgctcg gtcttgcctt gctcgtcggt
     5941 gatgtacaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
     6001 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
     6061 atagtttaaa ctgaaggcgg gaaacgacaa tctgatcccc ctccaccgcg gtggcggccg
     6121 cccggtcact ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa
     6181 atacaaatac atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataagaa
     6241 ccctaattcc cttatctggg aactactcac acattattct ggagaaaaat agagagagat
     6301 agatttgtag agagagactg gtgatttttg cggactctag agctcttact tgtacagctc
     6361 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     6421 cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag
     6481 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     6541 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     6601 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     6661 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     6721 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     6781 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     6841 gtagcgggcg aagcactgca cgccgtagcc gaaggtggtc acgagggtgg gccagggcac
     6901 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     6961 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     7021 gatgggcacc accccggtga acagctcctc gcccttgctc accatcccat caaccacttt
     7081 gtacaagaaa gctgggtcgg cgcgcccacc cttaggtgca ttcttcaatg aattttcggt
     7141 tatttcgata gccctgttgg tatactactg ccactcgttt atttgaagta aagacagcaa
     7201 aagttgttgc gctagatgac gaaggatgca catccagtac cttgtaagcg acaccgcgac
     7261 tcaaactagt gttttcatag acttcatcct ctatagtttt aaacggaaag tgttcaaagg
     7321 aattcatatt tgtatgtgaa taaacatctt cttcataaag tatgtcaccg cttcggcatt
     7381 gccacgttcg catggtgaag ggggcggccg cggagcctgc ttttttgtac aaacttgttg
     7441 atgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag ggatttgcag
     7501 aagtaacacc aaacaacagg gtgagcatcg acaaaagaaa cagtaccaag caaataaata
     7561 gcgtatgaag gcagggctaa aaaaatccac atatagctgc tgcatatgcc atcatccaag
     7621 t
//
