LOCUS       sequence-258909-        8960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CDS from the Zea mays ubiquitin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  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 8960)
  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:GW
FEATURES             Location/Qualifiers
     source          1..8960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1304..1483
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             1525..2076
                     /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      2096..2348
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2442..2466
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2778..3366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2858..2877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3537..4328,4329..4397))
                     /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     4160..4179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4398..4489)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4562..5271
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5383..5407
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(join(5700..6413,6414..6416,6417..6419))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5745..5766)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6094..6113
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6353..6374
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    6427..6551
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(6592..6897)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(6694..6713)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(7239..7898)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7813..7832
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7952..7958,7959..7976,7977..7982))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(8007..8131)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(8165..8181)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8166..8182)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgtctgcgga acggctagag ccatcccagg attccccaaa gagaaacact ggcaagttag
       61 caatcagaac gtgtctgacg tacaggtcgc atccgtgtac gaacgctagc agcacggatc
      121 tatcacaaac acggatctaa cacaaacatg aacagaagta gaactaccgg gccctaacca
      181 tggaccggaa cgccgatcta gagaaggtag agaggggggg gggaggacga gcggcgtacc
      241 ttgaagcgga ggtgccgacg ggtggatttg ggggagatct ggttgtgtgt gtgtgcgctc
      301 cgaacaacac gaggttgggg aaagagggtg tggagggggt gtctatttat tacggcgggc
      361 gaggaaggga aagcgaagga gcggtgggaa aggaatcccc cgtagctgcc ggtgccgtga
      421 gaggaggagg aggccgcctg ccgtgccggc tcacgtctgc cgctccgcca cgcaatttct
      481 ggatgccgac agcggagcaa gtccaacggt ggagcggaac tctcgagagg ggcccagagg
      541 cagcgacaga gatgccgtgc cgtctgcttc gcttggcccg acgcgacgct gctggttcgc
      601 tggttggtgt ccgttagact cgtcgacggc gtttaacagg ctggcattat ctactcgaaa
      661 caagaaaaat gtttccttag tttttttaat ttcttaaagg gtatttgttt aatttttagt
      721 cactttattt tattctattt tatatctaaa ttattaaata aaaaaactaa aatagagttt
      781 tagttttctt aatttagagg ctaaaataga ataaaataga tgtactaaaa aaattagtct
      841 ataaaaacca ttaaccctaa accctaaatg gatgtactaa taaaatggat gaagtattat
      901 ataggtgaag ctatttgcaa aaaaaaagga gaacacatgc acactaaaga gataaaactg
      961 tagagtcctg ttgtcaaagt actcaattgt cctttagacc atgtctaact gttcatttat
     1021 atgattctct aaaacactga tattattgta gtactataga ttatattatt cgtagagtga
     1081 agtttaaata tatgtataaa gatagataaa ctgcacttca aacaagtgtg acaaaaaaat
     1141 atgtggtaat tttttataac ttagacatgc aatgctcatt aaatcggcgc gcccaaaccg
     1201 aaggcgggaa acgacaatct gatcatgagc ggagaattaa gggagtcacg ttatgacccc
     1261 cgccgatgac gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag
     1321 gagccactga gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat
     1381 tattgcgcgt tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa
     1441 atgctccact gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc
     1501 tcaactcgat cgaggggatc taccatgagc ccagaacgac gcccggccga catccgccgt
     1561 gccaccgagg cggacatgcc ggcggtctgc accatcgtca accactacat cgagacaagc
     1621 acggtcaact tccgtaccga gccgcaggaa ccgcaggagt ggacggacga cctcgtccgt
     1681 ctgcgggagc gctatccctg gctcgtcgcc gaggtggacg gcgaggtcgc cggcatcgcc
     1741 tacgcgggtc cctggaaggc acgcaacgcc tacgactgga cggccgagtc gaccgtgtac
     1801 gtctcccccc gccaccagcg gacgggactg ggctccacgc tctacaccca cctgctgaag
     1861 tccctggagg cacagggctt caagagcgtg gtcgctgtca tcgggctgcc caacgacccg
     1921 agcgtgcgca tgcacgaggc gctcggatat gccccccgcg gcatgctgcg ggcggccggc
     1981 ttcaagcacg ggaactggca tgacgtgggt ttctggcagc tggacttcag cctgccggtg
     2041 ccgccccgtc cggtcctgcc cgtcaccgaa atctgatgac ccctagagtc aagcagatcg
     2101 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     2161 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     2221 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     2281 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     2341 actagatcga ccggcatgca agctgatatc tggtagagac gttagtattc gatcattaaa
     2401 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
     2461 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
     2521 acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag gcggcagact
     2581 ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg tttgaaacac
     2641 ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt gctgcctgtg
     2701 atcctctagc tagagtccat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2761 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2821 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2881 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2941 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3001 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3061 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3121 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3181 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3241 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3301 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtagct
     4501 ttcgggctga tcgcccatcc cgttactgcc ccgatcccgg caatggcaag gactgccagc
     4561 gctgccattt ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg
     4621 aggcccgcgt tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg
     4681 cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa
     4741 agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct
     4801 ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc
     4861 actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa
     4921 gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc
     4981 gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa
     5041 tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac
     5101 gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg
     5161 gccggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
     5221 ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cggaaaggcg ctcggtcttg
     5281 ccttgctcgt cggtgatgta cacatacaaa tggacgaacg gataaacctt ttcacgccct
     5341 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
     5401 cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat ccccctccac
     5461 cgcggtggcg gccgcccggt cactggattt tggttttagg aattagaaat tttattgata
     5521 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
     5581 aaaccctata agaaccctaa ttcccttatc tgggaactac tcacacatta ttctggagaa
     5641 aaatagagag agatagattt gtagagagag actggtgatt tttgcggact ctagagctct
     5701 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     5761 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag
     5821 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     5881 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     5941 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     6001 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     6061 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     6121 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     6181 ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agccgaaggt ggtcacgagg
     6241 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     6301 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     6361 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatc
     6421 ccatcaacca ctttgtacaa gaaagctgaa cgagaaacgt aaaatgatat aaatatcaat
     6481 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     6541 cagtcactat ggtcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc
     6601 ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac
     6661 ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct
     6721 ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg
     6781 tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac
     6841 atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc
     6901 accagcccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag
     6961 ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct
     7021 gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata
     7081 gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata
     7141 cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca
     7201 tactgttatc tggcttttag taagccggat ccagatcttt acgccccgcc ctgccactca
     7261 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     7321 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     7381 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     7441 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     7501 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     7561 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     7621 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg
     7681 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     7741 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     7801 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     7861 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     7921 aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc ataaagtgta
     7981 aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt ttacagtatt
     8041 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     8101 tttctcgttc agcttttttg tacaaacttg ttgatgggct gcaggaattc gatatcaagc
     8161 ttatcgatac cgtcgacctc gagggatttg cagaagtaac accaaacaac agggtgagca
     8221 tcgacaaaag aaacagtacc aagcaaataa atagcgtatg aaggcagggc taaaaaaatc
     8281 cacatatagc tgctgcatat gccatcatcc aagtatatca agatcaaaat aattataaaa
     8341 catacttgtt tattataata gataggtact caaggttaga gcatatgaat agatgctgca
     8401 tatgccatca tgtatatgca tcagtaaaac ccacatcaac atgtatacct atcctagatc
     8461 gatacttcca tccatcatct taaactcgta actatgaaga tgtatggcac acacatacag
     8521 atccaaaatt aataaatcca ccagttagtt tgaaacagta ttctactccg atctagaacg
     8581 aatgaacgac cgcccaacca caccacatca tcacaaccaa gcgaacaaaa agcatctctg
     8641 tatatgcatc agtaaaaccc gcatcaacat gtatacctat cctagatcga tatttccatc
     8701 catcatcttc aattcgtaac tatgaatatg tatggcacac acatacagat ccaaaattaa
     8761 taaatccacc aggtagtttg aaacagaatt ctactccgat ttagaacgac cgcccaacca
     8821 gaccacatca tcacaaccaa gccaaaaaaa aaacatgaaa agatgacccg acaaacaagt
     8881 gcacggcata tattgaaata aaggaaaagg gcaaaccaaa ccctatgcaa cgaaacaaaa
     8941 aaaaatcatg aaatcgatcc
//
