LOCUS       sequence-274750-        6381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luminescent reporter for cytokinin signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6381)
  AUTHORS   Lehmann S, Dominguez-Ferreras A, Huang WJ, Denby K, Ntoukakis V, 
            Schafer P
  TITLE     Novel markers for high-throughput protoplast-based analyses of 
            phytohormone signaling.
  JOURNAL   PLoS One. 2020 Jun 4;15(6):e0234154. doi: 
            10.1371/journal.pone.0234154. eCollection 2020.
  PUBMED    32497144
REFERENCE   2  (bases 1 to 6381)
  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: "PLoS One.";
            date: "2020-06-4"; volume: "15(6):e0234154. doi"; pages: " 
            10.1371/journal.pone.0234154. eCollection 2020"
COMMENT     Sequence Label: pARR5::LUC
FEATURES             Location/Qualifiers
     source          1..6381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..31)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15..31)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(28..50)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    39..55
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(63..69,70..87,88..93))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(246..263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(417..1005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(497..516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1176..1967,1968..2036))
                     /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     1799..1818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2037..2141)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2209..2227
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2387..2406
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2600..2622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2614..2631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2615..2631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4318..5970
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4384..4402)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      6125..6376
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
       61 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      121 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      181 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      841 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2161 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2221 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2281 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2341 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     2401 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2461 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     2521 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     2581 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
     2641 gcatgcctgc aggctctaga ggatccccaa taaagcatat ttgtcttcag cctctcatac
     2701 tatttgagag gtaaaaaccg agaccattag gattattctt tatagaatgt tttggtgcct
     2761 aacaaatttt actggcattt cttttttttg ccataaactg ttaagaggtt tatttatgaa
     2821 ttgttaactt aatttctttg atcaaacaat agtaagttct ataaatgtat cttgtgatct
     2881 ttgactatga tagacactaa gctcggactc atgtacatag ccaaaacact catatatatt
     2941 aaaacaataa tttcatcggt aaactcatgt tgtttccaaa tatacaagta gagaaaacgt
     3001 gtagacttac tctgaaggga cacgcatata catactgata aacaaccatc atggagaaat
     3061 ccgagccgtt tctctgcaag ccaaaccaaa tttgacatat atagtaaggt catgtgctct
     3121 atacacctag tacctcacat tcatttgata tctcctctag agatacatcg atcatagtca
     3181 tattttcagc tttcaatgat gatttagtca ttaccattct tgaaatatat tggtcgagtt
     3241 tagtccatag atcttcgatc ctgtcgtgac aaagccttcc atcaattgtc aaaagatctc
     3301 tcacgtgtgg aaatactcga tgcgtgttgc taaatatgtg aataaaagaa cgccaaattt
     3361 atagaaaaag cgaattaaaa ttcaagtaaa catatgaata tttttcaaga aaagtgaatt
     3421 gaaattttat tttcatgaaa acttactatt tttttttttt tttttgaaca ctcatgaaaa
     3481 cttactatta actcatcaat cttatgccat tcgaccacga tcgatagtag aggcaactat
     3541 tatggtacag ttacgagatg gacaaacaac aataacttga aaaataatga ttttttgttt
     3601 tgtttctaga aatatacaca tacggatctt gaatttatct gattgcgcaa ctagagatat
     3661 caacattgaa attcattcgc ccaaatagtc aatctggcgg tcacatgtca gctaactttt
     3721 ccaacaatgt tcattaatta attaacaatc tcaaagattt tgtagattga aatacaaatc
     3781 ttctctctgt ggtacatttc ttgaaaaatg ggaaaatcaa gaaagtatcg aaaatgtaca
     3841 aaaataaaaa gaaatgaatc aaagtagcca tgatcttgac aacaataatc gagagagatc
     3901 gtcatgatac gatttccctc atccaaaatt gattttattt cccttcccaa atcaaacata
     3961 tcatatgatt tcaccactca ccattacttg actattctca acaaaaaaaa tattaaaaaa
     4021 ctttatgact ttgattttat ttttatttga agtttagcca aaatttgaaa atatgacttt
     4081 tgagaagaaa acagaataaa caaataatta gccacgcgct atcagacaga caaaatccca
     4141 cagatatgca aagatctctc agaatcctct ccccatatca tatttttctc ttttccctct
     4201 ccttctttct tcctttataa atccatttat tctcctctca tctctcagca aaatcaaatc
     4261 ctcatagttg attctctcta tctctctcac gagtcacgat cctactcttc ttgatccatg
     4321 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atcctctaga ggatggaacc
     4381 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     4441 tttacagatg cacatatcga ggtgaacatc acgtacgcgg aatacttcga aatgtccgtt
     4501 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     4561 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     4621 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gaacatttcg
     4681 cagcctaccg tagtgtttgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     4741 aaattaccaa taatccagaa aattattatc atggattcta aaacggatta ccagggattt
     4801 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     4861 gtaccagagt cctttgatcg tgacaaaaca attgcactga taatgaattc ctctggatct
     4921 actgggttac ctaagggtgt ggcccttccg catagaactg cctgcgtcag attctcgcat
     4981 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     5041 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     5101 gtcgtcttaa tgtatagatt tgaagaagag ctgtttttac gatcccttca ggattacaaa
     5161 attcaaagtg cgttgctagt accaacccta ttttcattct tcgccaaaag cactctgatt
     5221 gacaaatacg atttatctaa tttacacgaa attgcttctg ggggcgcacc tctttcgaaa
     5281 gaagtcgggg aagcggttgc aaaacgcttc catcttccag ggatacgaca aggatatggg
     5341 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     5401 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     5461 ctgggcgtta atcagagagg cgaattatgt gtcagaggac ctatgattat gtccggttat
     5521 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     5581 gacatagctt actgggacga agacgaacac ttcttcatag ttgaccgctt gaagtcttta
     5641 attaaataca aaggatatca ggtggccccc gctgaattgg aatcgatatt gttacaacac
     5701 cccaacatct tcgacgcggg cgtggcaggt cttcccgacg atgacgccgg tgaacttccc
     5761 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     5821 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     5881 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     5941 gccaagaagg gcggaaagtc caaattgtaa aatgtaactg tattcagcga tgacgaaatt
     6001 cttagctatt gtaatattat atgcaaattg atgaatggta attttgtaat tgtgggtcac
     6061 tgtactattt taacgaataa taaaatcagg tataggtaac taaaaaggaa ttcgagctcg
     6121 aattgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     6181 tgcgatgatt atcatataat ttctgttgat tacgttaagc atgtaataat taacatgtaa
     6241 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     6301 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     6361 tctatgttac tagatctcta g
//
