LOCUS       sequence-258863-        5997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of firefly luciferase from the Zea mays 
            ubiquitin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5997)
  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 5997)
  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:LUC
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        283..329
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     regulatory      342..351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             348..2000
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(414..432)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      2046..2298
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2343..2447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2448..2516,2517..3308)
                     /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(2666..2685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3479..4067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3968..3987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4221..4238
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtatcgatc taggataggt atacatgttg atgtgggttt tactgatgca tatacatgat
       61 ggcatatgca gcatctattc atatgctcta accttgagta cctatctatt ataataaaca
      121 agtatgtttt ataattattt tgatcttgat atacttggat gatggcatat gcagcagcta
      181 tatgtggatt tttttagccc tgccttcata cgctatttat ttgcttggta ctgtttcttt
      241 tgtcgatgct cactagtgcc ggcctgcagt ctagagtcga ccgcaagacc cttcctctat
      301 ataaggaagt tcatttcatt tggagaggac acgctcgagt ggccaccatg gaagacgcca
      361 aaaacataaa gaaaggcccg gcgccattct atcctcttga ggatggaacc gctggagagc
      421 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
      481 cacatatcga ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt cggttggcag
      541 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
      601 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
      661 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gaacatttcg cagcctaccg
      721 tagtgtttgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aaattaccaa
      781 taatccagaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
      841 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtaccagagt
      901 cctttgatcg tgacaaaaca attgcactga taatgaactc ctctggatct actgggttac
      961 ctaagggtgt ggcccttccg catagaactg cctgcgtcag attctcgcat gccagagatc
     1021 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     1081 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     1141 tgtatagatt tgaagaagag ctgtttttac gatcccttca ggattacaaa attcaaagtg
     1201 cgttgctagt accaacccta ttttcattct tcgccaaaag cactctgatt gacaaatacg
     1261 atttatctaa tttacacgaa attgcttctg ggggcgcacc tctttcgaaa gaagtcgggg
     1321 aagcggttgc aaaacgcttc catcttccag ggatacgaca aggatatggg ctcactgaga
     1381 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     1441 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     1501 atcagagagg cgaattatgt gtcagaggac ctatgattat gtccggttat gtaaacaatc
     1561 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     1621 actgggacga agacgaacac ttcttcatag ttgaccgctt gaagtcttta attaaataca
     1681 aaggatatca ggtggccccc gctgaattgg aatcaatatt gttacaacac cccaacatct
     1741 tcgacgcggg cgtggcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     1801 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     1861 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     1921 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     1981 gcggaaagtc caaattgtaa gagctcccgg gtacctgatc atgagtaatt agctcgaatt
     2041 tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
     2101 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
     2161 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
     2221 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
     2281 catctatgtt actagatcgg gaattagatc tgctagcagg tggcactttt cggggaaatg
     2341 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2401 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2461 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2521 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2581 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2641 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2701 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2761 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2821 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2881 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2941 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3001 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3061 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4141 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4201 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagtggatca agcttggatc
     4321 cttaatgagc attgcatgtc taagttataa aaaattacca catatttttt tgtcacactt
     4381 gtttgaagtg cagtttatct atctttatac atatatttaa acttcactct acgaataata
     4441 taatctatag tactacaata atatcagtgt tttagagaat catataaatg aacagttaga
     4501 catggtctaa aggacaattg agtactttga caacaggact ctacagtttt atctctttag
     4561 tgtgcatgtg ttctcctttt tttttgcaaa tagcttcacc tatataatac ttcatccatt
     4621 ttattagtac atccatttag ggtttagggt taatggtttt tatagactaa tttttttagt
     4681 acatctattt tattctattt tagcctctaa attaagaaaa ctaaaactct attttagttt
     4741 ttttatttaa taatttagat ataaaataga ataaaataaa gtgactaaaa attaaacaaa
     4801 taccctttaa gaaattaaaa aaactaagga aacatttttc ttgtttcgag tagataatgc
     4861 cagcctgtta aacgccgtcg acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg
     4921 cgtcgggcca agcgaagcag acggcacggc atctctgtcg ctgcctctgg gcccctctcg
     4981 agagttccgc tccaccgttg gacttgctcc gctgtcggca tccagaaatt gcgtggcgga
     5041 gcggcagacg tgagccggca cggcaggcgg cctcctcctc ctctcacggc accggcagct
     5101 acgggggatt cctttcccac cgctccttcg ctttcccttc ctcgcccgcc gtaataaata
     5161 gacaccccct ccacaccctc tttccccaac ctcgtgttgt tcggagcgca cacacacaca
     5221 accagatctc ccccaaatcc acccgtcggc acctccgctt caaggtacgc cgctcgtcct
     5281 cccccccccc tctctacctt ctctagatcg gcgttccggt ccatggttag ggcccggtag
     5341 ttctacttct gttcatgttt gtgttagatc cgtgtttgtg atagatccgt gctgctagcg
     5401 ttcgtacacg gatgcgacct gtacgtcaga cacgttctga ttgctaactt gccagtgttt
     5461 ctctttgggg aatcctggga tggctctagc cgttccgcag acgggatcga tttcatgatt
     5521 tttttttgtt tcgttgcata gggtttggtt tgcccttttc ctttatttca atatatgccg
     5581 tgcacttgtt tgtcgggtca tcttttcatg tttttttttt ggcttggttg tgatgatgtg
     5641 gtctggttgg gcggtcgttc taaatcggag tagaattctg tttcaaacta cctggtggat
     5701 ttattaattt tggatctgta tgtgtgtgcc atacatattc atagttacga attgaagatg
     5761 atggatggaa atatcgatct aggataggta tacatgttga tgcgggtttt actgatgcat
     5821 atacagagat gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg gcggtcgttc
     5881 attcgttcta gatcggagta gaatactgtt tcaaactaac tggtggattt attaattttg
     5941 gatctgtatg tgtgtgccat acatcttcat agttacgagt ttaagatgat ggatgga
//
