LOCUS       sequence-254163-        5997 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Transient expression of firefly luciferase from the Zea mays 
            ubiquitin promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
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="genomic DNA"
                     /organism="unspecified"
     promoter        1959..2005
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     regulatory      2018..2027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2024..3676
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2090..2108)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      3722..3974
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        4019..4123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4124..4192,4193..4984)
                     /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(4342..4361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5155..5743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5644..5663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5897..5914
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttaatgagca ttgcatgtct aagttataaa aaattaccac atattttttt gtcacacttg
       61 tttgaagtgc agtttatcta tctttataca tatatttaaa cttcactcta cgaataatat
      121 aatctatagt actacaataa tatcagtgtt ttagagaatc atataaatga acagttagac
      181 atggtctaaa ggacaattga gtactttgac aacaggactc tacagtttta tctctttagt
      241 gtgcatgtgt tctccttttt ttttgcaaat agcttcacct atataatact tcatccattt
      301 tattagtaca tccatttagg gtttagggtt aatggttttt atagactaat ttttttagta
      361 catctatttt attctatttt agcctctaaa ttaagaaaac taaaactcta ttttagtttt
      421 tttatttaat aatttagata taaaatagaa taaaataaag tgactaaaaa ttaaacaaat
      481 accctttaag aaattaaaaa aactaaggaa acatttttct tgtttcgagt agataatgcc
      541 agcctgttaa acgccgtcga cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc
      601 gtcgggccaa gcgaagcaga cggcacggca tctctgtcgc tgcctctggg cccctctcga
      661 gagttccgct ccaccgttgg acttgctccg ctgtcggcat ccagaaattg cgtggcggag
      721 cggcagacgt gagccggcac ggcaggcggc ctcctcctcc tctcacggca ccggcagcta
      781 cgggggattc ctttcccacc gctccttcgc tttcccttcc tcgcccgccg taataaatag
      841 acaccccctc cacaccctct ttccccaacc tcgtgttgtt cggagcgcac acacacacaa
      901 ccagatctcc cccaaatcca cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc
      961 ccccccccct ctctaccttc tctagatcgg cgttccggtc catggttagg gcccggtagt
     1021 tctacttctg ttcatgtttg tgttagatcc gtgtttgtga tagatccgtg ctgctagcgt
     1081 tcgtacacgg atgcgacctg tacgtcagac acgttctgat tgctaacttg ccagtgtttc
     1141 tctttgggga atcctgggat ggctctagcc gttccgcaga cgggatcgat ttcatgattt
     1201 ttttttgttt cgttgcatag ggtttggttt gcccttttcc tttatttcaa tatatgccgt
     1261 gcacttgttt gtcgggtcat cttttcatgt tttttttttg gcttggttgt gatgatgtgg
     1321 tctggttggg cggtcgttct aaatcggagt agaattctgt ttcaaactac ctggtggatt
     1381 tattaatttt ggatctgtat gtgtgtgcca tacatattca tagttacgaa ttgaagatga
     1441 tggatggaaa tatcgatcta ggataggtat acatgttgat gcgggtttta ctgatgcata
     1501 tacagagatg ctttttgttc gcttggttgt gatgatgtgg tgtggttggg cggtcgttca
     1561 ttcgttctag atcggagtag aatactgttt caaactaact ggtggattta ttaattttgg
     1621 atctgtatgt gtgtgccata catcttcata gttacgagtt taagatgatg gatggaagta
     1681 tcgatctagg ataggtatac atgttgatgt gggttttact gatgcatata catgatggca
     1741 tatgcagcat ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta
     1801 tgttttataa ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg
     1861 tggatttttt tagccctgcc ttcatacgct atttatttgc ttggtactgt ttcttttgtc
     1921 gatgctcact agtgccggcc tgcagtctag agtcgaccgc aagacccttc ctctatataa
     1981 ggaagttcat ttcatttgga gaggacacgc tcgagtggcc accatggaag acgccaaaaa
     2041 cataaagaaa ggcccggcgc cattctatcc tcttgaggat ggaaccgctg gagagcaact
     2101 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     2161 tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt tggcagaagc
     2221 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     2281 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     2341 cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc ctaccgtagt
     2401 gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaaat taccaataat
     2461 ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     2521 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac cagagtcctt
     2581 tgatcgtgac aaaacaattg cactgataat gaactcctct ggatctactg ggttacctaa
     2641 gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca gagatcctat
     2701 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     2761 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     2821 tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt
     2881 gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca aatacgattt
     2941 atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag tcggggaagc
     3001 ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca ctgagactac
     3061 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     3121 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     3181 gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa acaatccgga
     3241 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     3301 ggacgaagac gaacacttct tcatagttga ccgcttgaag tctttaatta aatacaaagg
     3361 atatcaggtg gcccccgctg aattggaatc aatattgtta caacacccca acatcttcga
     3421 cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     3481 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     3541 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     3601 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     3661 aaagtccaaa ttgtaagagc tcccgggtac ctgatcatga gtaattagct cgaatttccc
     3721 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     3781 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     3841 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     3901 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     3961 tatgttacta gatcgggaat tagatctgct agcaggtggc acttttcggg gaaatgtgcg
     4021 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4081 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4501 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4681 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4801 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4981 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5821 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5881 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5941 accgcctctc cccgcgcgtt ggccgattca ttaatgcagt ggatcaagct tggatcc
//
