LOCUS       sequence-258865-        7623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Blumeria graminis f. sp. hordei effector AVRa1 ∆SP
            from the Zea mays ubiquitin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7623)
  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 7623)
  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
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(156..869,870..872,873..875))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(201..222)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     550..569
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     809..830
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    883..907
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1226..1250)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1284..1300)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1285..1301)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        3383..3562
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             3604..4155
                     /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      4175..4427
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4521..4545
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4857..5445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4937..4956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5616..6407,6408..6476))
                     /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     6239..6258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6477..6568)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6641..7350
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    7462..7486
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataagaa ccctaattcc
       61 cttatctggg aactactcac acattattct ggagaaaaat agagagagat agatttgtag
      121 agagagactg gtgatttttg cggactctag agctcttact tgtacagctc gtccatgccg
      181 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
      241 gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
      301 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
      361 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
      421 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
      481 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
      541 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
      601 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
      661 aagcactgca cgccgtagcc gaaggtggtc acgagggtgg gccagggcac gggcagcttg
      721 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
      781 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
      841 accccggtga acagctcctc gcccttgctc accatcccat caaccacttt gtacaagaaa
      901 gctgggtcgg cgcgcccacc cttctaggtg cattcttcaa tgaattttcg gttatttcga
      961 tagccctgtt ggtatactac tgccactcgt ttatttgaag taaagacagc aaaagttgtt
     1021 gcgctagatg acgaaggatg cacatccaat accttgtaaa cgacaccgcg actcaaacta
     1081 gtgttttcat agacttcatc ctctatagtt ttaaacggaa agtgttcaaa ggaattcata
     1141 tttgtatgtg aataaacatc ttcttcataa agtatgtcac cgcttgggca ttgccacgtt
     1201 cgcatggtga agggggcggc cgcggagcct gcttttttgt acaaacttgt tgatgggctg
     1261 caggaattcg atatcaagct tatcgatacc gtcgacctcg agggatttgc agaagtaaca
     1321 ccaaacaaca gggtgagcat cgacaaaaga aacagtacca agcaaataaa tagcgtatga
     1381 aggcagggct aaaaaaatcc acatatagct gctgcatatg ccatcatcca agtatatcaa
     1441 gatcaaaata attataaaac atacttgttt attataatag ataggtactc aaggttagag
     1501 catatgaata gatgctgcat atgccatcat gtatatgcat cagtaaaacc cacatcaaca
     1561 tgtataccta tcctagatcg atacttccat ccatcatctt aaactcgtaa ctatgaagat
     1621 gtatggcaca cacatacaga tccaaaatta ataaatccac cagttagttt gaaacagtat
     1681 tctactccga tctagaacga atgaacgacc gcccaaccac accacatcat cacaaccaag
     1741 cgaacaaaaa gcatctctgt atatgcatca gtaaaacccg catcaacatg tatacctatc
     1801 ctagatcgat atttccatcc atcatcttca attcgtaact atgaatatgt atggcacaca
     1861 catacagatc caaaattaat aaatccacca ggtagtttga aacagaattc tactccgatt
     1921 tagaacgacc gcccaaccag accacatcat cacaaccaag ccaaaaaaaa aacatgaaaa
     1981 gatgacccga caaacaagtg cacggcatat attgaaataa aggaaaaggg caaaccaaac
     2041 cctatgcaac gaaacaaaaa aaaatcatga aatcgatccc gtctgcggaa cggctagagc
     2101 catcccagga ttccccaaag agaaacactg gcaagttagc aatcagaacg tgtctgacgt
     2161 acaggtcgca tccgtgtacg aacgctagca gcacggatct atcacaaaca cggatctaac
     2221 acaaacatga acagaagtag aactaccggg ccctaaccat ggaccggaac gccgatctag
     2281 agaaggtaga gagggggggg ggaggacgag cggcgtacct tgaagcggag gtgccgacgg
     2341 gtggatttgg gggagatctg gttgtgtgtg tgtgcgctcc gaacaacacg aggttgggga
     2401 aagagggtgt ggagggggtg tctatttatt acggcgggcg aggaagggaa agcgaaggag
     2461 cggtgggaaa ggaatccccc gtagctgccg gtgccgtgag aggaggagga ggccgcctgc
     2521 cgtgccggct cacgtctgcc gctccgccac gcaatttctg gatgccgaca gcggagcaag
     2581 tccaacggtg gagcggaact ctcgagaggg gcccagaggc agcgacagag atgccgtgcc
     2641 gtctgcttcg cttggcccga cgcgacgctg ctggttcgct ggttggtgtc cgttagactc
     2701 gtcgacggcg tttaacaggc tggcattatc tactcgaaac aagaaaaatg tttccttagt
     2761 ttttttaatt tcttaaaggg tatttgttta atttttagtc actttatttt attctatttt
     2821 atatctaaat tattaaataa aaaaactaaa atagagtttt agttttctta atttagaggc
     2881 taaaatagaa taaaatagat gtactaaaaa aattagtcta taaaaaccat taaccctaaa
     2941 ccctaaatgg atgtactaat aaaatggatg aagtattata taggtgaagc tatttgcaaa
     3001 aaaaaaggag aacacatgca cactaaagag ataaaactgt agagtcctgt tgtcaaagta
     3061 ctcaattgtc ctttagacca tgtctaactg ttcatttata tgattctcta aaacactgat
     3121 attattgtag tactatagat tatattattc gtagagtgaa gtttaaatat atgtataaag
     3181 atagataaac tgcacttcaa acaagtgtga caaaaaaata tgtggtaatt ttttataact
     3241 tagacatgca atgctcatta aatcggcgcg cccaaaccga aggcgggaaa cgacaatctg
     3301 atcatgagcg gagaattaag ggagtcacgt tatgaccccc gccgatgacg cgggacaagc
     3361 cgttttacgt ttggaactga cagaaccgca acgttgaagg agccactgag ccgcgggttt
     3421 ctggagttta atgagctaag cacatacgtc agaaaccatt attgcgcgtt caaaagtcgc
     3481 ctaaggtcac tatcagctag caaatatttc ttgtcaaaaa tgctccactg acgttccata
     3541 aattcccctc ggtatccaat tagagtctca tattcactct caactcgatc gaggggatct
     3601 accatgagcc cagaacgacg cccggccgac atccgccgtg ccaccgaggc ggacatgccg
     3661 gcggtctgca ccatcgtcaa ccactacatc gagacaagca cggtcaactt ccgtaccgag
     3721 ccgcaggaac cgcaggagtg gacggacgac ctcgtccgtc tgcgggagcg ctatccctgg
     3781 ctcgtcgccg aggtggacgg cgaggtcgcc ggcatcgcct acgcgggtcc ctggaaggca
     3841 cgcaacgcct acgactggac ggccgagtcg accgtgtacg tctccccccg ccaccagcgg
     3901 acgggactgg gctccacgct ctacacccac ctgctgaagt ccctggaggc acagggcttc
     3961 aagagcgtgg tcgctgtcat cgggctgccc aacgacccga gcgtgcgcat gcacgaggcg
     4021 ctcggatatg ccccccgcgg catgctgcgg gcggccggct tcaagcacgg gaactggcat
     4081 gacgtgggtt tctggcagct ggacttcagc ctgccggtgc cgccccgtcc ggtcctgccc
     4141 gtcaccgaaa tctgatgacc cctagagtca agcagatcgt tcaaacattt ggcaataaag
     4201 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     4261 ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt
     4321 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     4381 aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcgac cggcatgcaa
     4441 gctgatatct ggtagagacg ttagtattcg atcattaaaa acgtccgcaa tgtgttatta
     4501 agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag
     4561 ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc atcagtccgg
     4621 gacggcgtca gcgggagagc cgttgtaagg cggcagactt tgctcatgtt accgatgcta
     4681 ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
     4741 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcctctagct agagtccatg
     4801 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4861 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4921 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4981 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5041 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5101 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5161 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5221 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5281 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5341 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5401 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5461 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5521 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5581 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5641 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5701 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5761 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5821 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5881 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5941 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6001 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6061 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6121 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6181 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6241 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6301 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6361 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6421 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6481 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6541 tttgaatgta tttagaaaaa taaacaaata ggggtagctt tcgggctgat cgcccatccc
     6601 gttactgccc cgatcccggc aatggcaagg actgccagcg ctgccatttt tggggtgagg
     6661 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     6721 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     6781 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     6841 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     6901 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     6961 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     7021 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     7081 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     7141 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     7201 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccggtgtctc gcacacggct
     7261 tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca gcggcgaggg
     7321 caaccagccc ggtgagcgtc ggaaaggcgc tcggtcttgc cttgctcgtc ggtgatgtac
     7381 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta
     7441 ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt
     7501 aaactgaagg cgggaaacga caatctgatc cccctccacc gcggtggcgg ccgcccggtc
     7561 actggatttt ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa
     7621 tac
//
